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Protests have erupted across the nation and returned to St. Louis: Here are some of the leaders of the movement – STLtoday.com

The Rev. Darryl Gray sits at the intersection of Market Street and Tucker Boulevard for minutes of silence in memory of those killed by police officers, near City Hall in downtown St. Louis on Wednesday, July 8, 2020. Protesters announced plans to camp in front of the building until their demands were met and Mayor Lyda Krewson resigns. Photo by Robert Cohen, rcohen@post-dispatch.com

Protests dont just happen as spontaneous moments.

They are planned and executed by an array of people college students and politicians, lawyers and social workers, medical professionals and blue-collar workers spurred by outrage, activism or a sense of injustice.

Protesters who emerged as leaders six years ago after 18-year-old Michael Brown was shot and killed by a Ferguson police officer have in some cases carried their commitment in new directions of influence, including freshman state Rep. Rasheen Aldridge and Cori Bush, the Democratic nominee in the 1st Congressional District after Tuesdays primary election.

Aldridge and Bush have remained active in protests, including those sparked this summer by the death of George Floyd, a Black man killed while restrained by a Minneapolis police officer, as well as by the release of a video that showed a Florissant police detective hitting a man with his vehicle.

But they are not the only leaders of the movement.

At the center of the demonstrations are a mix of groups ExpectUs, RespectUs, Tent Mission STL, Occupy City Hall STL, Protest THAT, Action St. Louis and ClosetheWorkhouse whose opinions vary on the best way to seek justice.

And on the anniversary of Browns death, Aug. 9, 2014, protest leaders say underlying inequities and a lack of broad reform measures make the St. Louis area ripe for social unrest.

Meet a few of the leaders who carry the culture of protest in the St. Louis region.

The pastor

The Rev. Daryl Gray, 66, is a founder of the protest group ExpectUs. He is one of several Democratic leaders in the organization who doubles as both an activist and a politician. Others include Aldridge and Bush, who toppled longtime U.S. Rep. William Lacy Clay in Tuesdays Democratic primary.

Among other calls for justice, ExpectUs has advocated for reparations for Black people and an end to police brutality. Gray said current protests are more of a movement than a moment.

Rev. Darryl Gray talks to police officers as protesters march at the Galleria in Richmond Heights on Black Friday, Nov. 24, 2017. About 100 people marched through the mall, calling for an economic boycott of area businesses. Photo by Robert Cohen, rcohen@post-dispatch.com

Born in Boston and having grown up in both South Carolina and Canada, Gray moved to St. Louis after Fergusons uprisings. He emphasizes that when an organization like ExpectUs is grassroots, its bound to have more buy-in from its members.

If you own it, then youre going to go the extra mile for it, Gray said.

Gray attended a newly integrated high school in South Carolina, where he got a taste of standing up for what he believed in and the repercussions that often follow.

During a school pep rally, Gray said a man came riding into the gym on a horse waving a Confederate flag. He walked out, and hundreds of students followed him. He was suspended from school, he said, for inciting a disturbance.

I didnt know they were behind me. I didnt look back, he said.

Gray went on to join the U.S. Army, where he was honorably discharged. He then campaigned for presidential candidate the Rev. Jesse Jackson in Florida, worked with the Southern Christian Leadership Conference, preached in both Canada and Atlanta, served as head of Atlantas NAACP, and briefly served as a state senator from Kansas City.

Now, he said, hes ready to make St. Louis his final destination.

With a history of frequently moving, proving himself in new cities has been the norm for Gray.

I think Ive gotten beat up enough in the streets. Ive shown up more than some people whove been here all their lives thats got to count for something, Gray said.

On July 6, a day after protesters say they were beaten while being arrested for protesting at Florissants police department, Gray asked officers in riot gear behind the gate of the police department to go back inside the police department as a sign of good faith. A white shirt officer agreed and instructed riot police to go inside, but a regular, uniformed group of police then came out another door.

Gray and other ExpectUs leaders told everyone to go home. Some in the crowd strongly opposed leaving though, and the two factions argued with one another.

That was our mistake the timing. For us to disperse while police were standing there could be perceived as a sign of weakness, Gray said. RespectUs felt disrespected at that moment, and they had reason to. Our timing was off. That was our mistake. We owned it.

The self-proclaimed radical

One of the protesters who stayed behind that night was Tauren Taylor, a 25-year-old University City resident.

A self-proclaimed radical, Taylor said he stayed because protesters were supposed to go out on their own terms. Taylor said protesters had to advocate for every inch of real estate on which to protest, including the parking lot across the street from the police station.

Tauren Taylor, center, talks with Dhoruba Shakur, left, and Tory Russell as they block the doorway to the Ferguson Market to shut it down for the night on Thursday, July 30, 2020, after St. Louis County prosecutor Wesley Bell announced that after a review he would not charge police officer Darren Wilson in Brown's 2014 death. The store became a central part of the case after the teenager was accused of robbing it before he was shot. Photo by Robert Cohen, rcohen@post-dispatch.com

Why keep letting them hand us stuff when our job is to take things? Were supposed to be forcing them to give us things they dont want to give us, Taylor said. They dont want to give us freedom. They dont want to stop killing us, they dont want to stop beating us.

Taylor has been arrested at several protests in the past year. He was also one of the protest leaders after Terry Tillman, 23, was killed last year by police near the St. Louis Galleria Mall in Richmond Heights.

Taylor remains vocal on the front line, but he says he will not join a protest group.

To be held under a name, youre held under their standards. ExpectUs, for instance, I love people in ExpectUS. I will ride for ExpectUs, but I dont think now is the time for us to try to ask for reparations.

Taylor moved to Missouri from California when he was a teenager. He said his family dealt with poverty, and he was bullied frequently for having a lisp and being studious.

One of his first successful protests, he said, was getting a neglectful teacher fired by collecting petition names.

Once I found out what the chain of the command was, I went up that ladder, he said.

Eventually, he said, he was kicked out of Vashon High School for fighting.

Taylor earned a diploma from the Fresh Start Academy program at 17, and briefly studied animal science and genetic engineering at St. Louis Community College at Forest Park.

He acknowledged some may see his protest methods as extreme, but he said his methods also help people understand someone is sticking up for them.

Taylor said hes not sure about a future career path, but hes working odd jobs, including selling homemade goods at Soulard Market. In the meantime, he said, he will continue to protest injustices wherever they spring up in the St. Louis area.

Tauren Taylor leads chants in the parking lot of the Florissant Police Department on Monday, July 6, 2020. Protesters held a demonstration against police brutality after a Florissant officer was fired and charged with hitting a fleeing subject with his vehicle. Those attending were seeking charges against two other officers involved in the incident. Photo by Robert Cohen, rcohen@post-dispatch.com

Taylor was one of several protesters to take exception with comments in mid-July by Jimmie Edwards, the top law enforcement official in St. Louis. Edwards, at an anti-crime demonstration outside City Hall, said many protesters werent from the city, and he condemned violence that erupted at some of the protests.

I feel like violence happens everywhere, even in nature, said Taylor. Were all human, we all make mistakes. Theres no way to control every single person thats out there. Ive seen police hurt people and each other. You cant condone it on one end and condemn it on the other.

The elder

The perceived lack of government response when doorbell security footage showed former Florissant Detective Joshua Smith running over a fleeing suspect in early June led to the formation of another protest group, RespectUs, said one of the founders, Cathy Mama Cat Daniels.

Daniels, 59, said she and a group of other front-line Ferguson protesters went to the police station for answers about what had happened, but there were barricades around the police department.

Protester Cathy Daniels leads the chant opposing the King Louis IX statue in Forest Park, during a rally atop Art Hill on Saturday, June 27, 2020. About 200 people on opposite sides of the debate attended the rally in front of the St. Louis Art Museum. Photo by Robert Cohen, rcohen@post-dispatch.com

When they didnt answer us and treat us with the human respect as people who live in this town, youve got to stand up and fight back, said Daniels. Even the (city) council, anyone who spoke against their idea of democracy, they didnt feel the need to respect them. Thats why we are RespectUs.

Daniels said RespectUs has four core demands: fire, charge, arrest and convict Smith. Thus far, three of those four have happened Smith has not yet had his day in court.

Daniels has lived in Florissant since 2012. She grew up in New York City and previously lived in Chicago and San Diego. Since Fergusons uprisings, shes worked as a cook and founded PotBangerz, a nonprofit dedicated to providing food and clothing to families in need. The organization is now renovating a home in Pine Lawn for cis, queer and trans women in need and plans to open the home later this year.

Cathy 'Mama Cat' Daniels empties one of two massive bowls of salad with vegetables donated from area farms, as she cooks dinner for 130 people in the kitchen at First Congregational Church of St. Louis on Thursday, July 23, 2020 in Clayton. Daniels leads the PotBangerz, a group of volunteers, many who are active in area protests, who feed the hungry and the unhoused. Photo by Robert Cohen, rcohen@post-dispatch.com

An elder of the protest group, Daniels said she offers advice to protesters and watches from the back, cane and chair in tow. Still, she said, she doesnt always approve of the protest methods that unfold.

Theres no such thing as peaceful protest. Thats an oxymoron. If there was peace, we wouldnt need to protest, Daniels said. I dont support tearing up stuff. We had that happen during Ferguson. If were gonna burn it down, then do it the right way, and thats not in the literal sense. Burn down the system. Defund the police.

The congresswoman

Daniels stood behind Cori Bush, 44, as hundreds of protesters returned July 3 to Portland Place in St. Louis West End. The private street had recently made national news, as Mark and Patricia McCloskey waved guns at protesters in an effort to, as they said, defend their home. The couple has since been charged with unlawful use of a weapon.

There was nothing to defend, and thats what angers me so bad, Bush said.

Protest leader Cori Bush marches on Forest Park Parkway during a rally and march against police brutality in the Delmar Loop on Friday, June 12, 2020. Photo by Robert Cohen, rcohen@post-dispatch.com

In addition to rising as a leader of ExpectUs, Bush gained national notoriety in Tuesdays election. In a district that has historically voted overwhelmingly for Democrats, Bush will likely become the first Black woman to represent Missouri in Congress.

Bush said she learned about politics, protest tactics and demonstrations from her father, Earl Bush, who worked as a politician in north St. Louis County.

She attended high school at Cardinal Ritter College Prep and said she had no intention of starting a life in politics. She graduated from Harris- Stowe State University and Lutheran School of Nursing, then entered the fields of nursing and ministry.

Between Bushs unsuccessful runs for Senate and the House of Representatives in 2016 and 2018, respectively, St. Louis police officer Jason Stockley went on trial for shooting a Black man, Anthony Lamar Smith, in 2011. Stockley was charged with first-degree murder, and prosecutors claimed he planted a gun on Smith. When Stockley was found not guilty by a judge in 2017, protests erupted again, and Bush emerged as a leader of ExpectUs.

During those demonstrations, Bush said, the group began to change how it disseminated information.

With Ferguson, you didnt have to call and ask around to see what time a protest was going on. You could just show up. It was 24/7. With Stockley, we had to have a way to get the information out, said Bush, highlighting the organizations use of social media.

Now, both she and Gray agree the groups protests are more refined, pointing out the local history lessons they provide before the marching and chanting begin.

ExpectUs doesnt do violent things, but what were not going to do is turn our backs on people that do, as far as their form of protest, Bush said, referencing the violence that occasionally breaks out at protests, including an early June demonstration that included gunfire and widespread looting as the night wore on.

If I have a sandwich today for breakfast, lunch and dinner Im not going to loot a sandwich, Bush said. If they fix the problems, they wont have to worry about that.

Bush said shes aware she could get blamed for the violence that occurs during protests, but she stands by her decision to continually show up and call for change.

I know it every single time, and I make that choice to show up, she said. When we stop showing up, when we stop pushing, thats how they win, so Ill take that chance.

This story has been updated to correct the year of the shooting of Anthony Lamar Smith.

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Protests have erupted across the nation and returned to St. Louis: Here are some of the leaders of the movement - STLtoday.com

A Unicorn Rivalry Shaping the Future of the Synbio Industry – SynBioBeta

Two companies Ginkgo Bioworks and Zymergen are poised to shape the future of SynBio

Across industries as diverse as electronics, pharmaceuticals, food, and materials, industrial synthetic biology has begun to produce the goods we need more efficiently and sustainably. Using the synthetic biology approachor synbio for those familiar with the fieldstart-ups and Fortune 500 corporations alike are genetically-engineering microbes to manufacture a massive range of goods from the touchscreens in your phone, to the cotton in your t-shirt, to the meat in your plant-based burger.

Traditionally, this synthetic biology process has been a slow and hypothesis-driven endeavor, dependent on researcher geniusand a healthy dose of luckto design these microbial mini-factories. But not anymore.

Two companies have embraced the power of computational biology to take the guesswork out of synthetic biology and drive this young industry into the mainstream. Zymergen and Ginkgo Bioworks both use vast metagenomic databases, machine learning, and robust automated laboratories to design microbes custom-suited to manufacture a desired good. Both organizations have raised significant capital to build out their platforms, with Zymergen having raised a $400-million Series C in 2018 and Ginkgo Bioworks raising almost double that figure in multiple rounds over its lifetime.

Despite their shared mission to turn the art of engineering biology into a science, the two synbio pioneers are strongly differentiated by their respective technology stacks and business models. Analyzing the key differences between the two can help us understand how the synthetic biology industry as a whole may evolve and where the value levers are hidden in the synbio ecosystem.

At its core, industrial synthetic biology offers a path to both producing humanitys typical array of materialssuch as plastics, textiles, food, and medicinesmore efficiently, as well as creating completely new substances. To produce these goods with biology relies on a three-step process of:

1) Molecule Identification: Identifying the molecule you want to create;

2) Biology Design: Designing a microbe to manufacture the good; and

3) End-Product Manufacturing: Scaling the manufacturing.

Overlaying this simplified three-step value chain against each of these organizations core competencies illustrates where the two companies hope to compete in the industrial synbio field and how they will likely co-exist (or not) with other players in the space.

Both Zymergen and Ginkgo have established wide competitive moats around the Biology Design process. By constructing massive and growing metagenomic databasesmapping genes, proteins, and metabolic pathwaysand investing capital into automated labs capable of running high-throughput experimentation, the two companies have achieved a level of take-off velocity insulating them from would-be challengers seeking to enter the field.

Both companies have embraced an R&D process that creates a positive feedback loop, perpetually self-improving their ability to design microbes and putting greater distance between them and any potential competitors. Both companies leverage their massive datasets to train machine-learning algorithms that help design potential microbes for a given purpose. They then turn to their automated labs to test thousands of strains for performance, creating new data-points to further improve their machine-learning for the next batch of microbes.

While important differences exist between the two companies Biology Design stacks regarding the size of their datasets, the robustness of their software, and the automation of their hardware, both can claim strong proficiency in the Biology Design portion of the synbio value chain.

Beyond Biology Design, Ginkgo and Zymergen have taken divergent approaches to both Molecule Identification and End-Product Manufacturing. Regarding the former, Zymergen has developed chemistry and materials datasets and supporting lab infrastructure to rigorously scan their databases for promising molecules, predict their fit for a given purpose, and test those predictions empirically. To its credit, Zymergen has announced three products so fara biofilm for electronics, an insect repellent, and a crop pest control agentthat are all reported to be completely novel, never-used-before molecules.

Zymergen has also endeavored to internalize End-Product Manufacturing capabilities. After Zymergen designs a product, they turn to a network of close bio-manufacturing partners to produce the end-product in-house for downstream customers and partners.

While exceptions exist, Ginkgo has concentrated its focus almost solely on the Biology Design pieceperhaps with good reason. The ability to predict how a new molecule will behave varies across applications and necessitates a diverse toolkit: the analysis of a molecule to be used in electronic consumables varies greatly from that of one to be used in food or nutrition, diluting the scalability of a Molecule Identification skillset. For this reason, Ginkgo has focused their platform less on the discovery of totally novel substances, but more on finding a better means of manufacturing those we already know.

In regard to End-Product Manufacturing, commercial bio-manufacturing represents a fairly old technologyit relies primarily on the same fermentation process used to make beer.

Accordingly, Ginkgo believes that the End-Product Manufacturing element of the synbio process is relatively commoditized, and that their platform does not provide much marginal advantage here. In Ginkgos worldview, Molecule Identification appears to be too specialized beyond their core competency, while End-Product Manufacturing is perhaps too commoditized to offer much additional value.

In a nutshell, Zymergen has positioned itself as a products company with the infrastructure to cover the full product development process, while Ginkgo acts as a platform providing a modular solution at one valuable link in the synbio value chain. Under this strategy, Zymergen can produce differentiated products that cannot be found anywhere elseand Zymergen will be able to charge for that unique value.

Lacking an internal products engine, Ginkgo has cast a wide net of partnerships with industry innovators and internal spin-outs, in effect creating a decentralized product discovery network that can leverage its biology design platform to bring products to market.

While the two companies early strategies may not be indicative of their future evolution, their current directions do reflect their intended role within the larger synthetic biology ecosystem. On the one hand, Zymergen seems to be driving for a world where they can act as the one-stop-shop for anyones manufacturing or materials needsthey can ID the best substance, engineer the best organisms to produce it, and manufacture the product for sale to customers.

Alternatively, Ginkgos platform allows for a more distributed ecosystemwhere they can leverage their unique expertise in organism design to provide downstream partners with the crucial key to implementing their own bio-manufacturing processes.

If either approach can succeed in pulling synthetic biology into the mainstream, we could all benefit from a cleaner, healthier, and more abundant world.

Matthew Kirshner works in life sciences consulting at Putnam Associates, focused on pharma and biotech organizations commercializing novel therapeutics and diagnostics. He has no professional affiliation in his work with either company referenced in this article.

Originally published on Genetic Engineering & Biotechnology News https://www.genengnews.com/gen-edge/a-unicorn-rivalry-shaping-the-future-of-the-synbio-industry/

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Discovery Could Lead to More Potent Garlic, Boosting Flavor and Aroma – SciTechDaily

Hannah Valentino, left, and Pablo Sobrado, right, uncover a new step in the process that makes garlic potent. Credit: Virginia Tech

For centuries, people around the world have used garlic as a spice, natural remedy, and pest deterrent but they didnt know how powerful or pungent the heads of garlic were until they tasted them.

This information changes the whole story about how garlic could be improved. Hannah Valentino, a Ph.D. candidate in the Virginia Tech College of Agriculture and Life Sciences

But what if farmers were able to grow garlic and know exactly how potent it would be? What if buyers could pick their garlic based on its might?

A team of Virginia Tech researchers recently discovered a new step in the metabolic process that produces the enzyme allicin, which leads to garlics delectable flavor and aroma, a finding that upends decades of previous scientific belief. Their work could boost the malodorous yet delicious characteristics that garlic-lovers the world over savor.

This information changes the whole story about how garlic could be improved or we could make the compounds responsible for its unique flavor, said Hannah Valentino, a College of Agriculture and Life Sciences Ph.D. candidate. This could lead to a new strain of garlic that would produce more flavor.

The discovery of this pathway opens the door for better control of production and more consistent crops, which would help farmers. Garlic could be sold as strong or weak, depending on consumer preferences.

The research was recently published in the Journal of Biological Chemistry.

Hannah Valentino, left, and Pablo Sobrado, right, are conducting research that is laying the foundation for a future in which buyers can choose garlic based on its strength and flavor profile. Credit: Virginia Tech

When Valentino, an Institute for Critical Technology and Applied Science doctoral fellow, and her team set out to test the generally accepted biological process that creates allicin, they found it just didnt happen.

Thats when the team of researchers set out to discover what was really happening in garlic.

As they peeled back the layers, they realized there was no fuel to power the previously accepted biological process that creates allicin.

By using rational design, Hannah found a potential substrate, said Pablo Sobrado, professor of biochemistry in the College of Agriculture and Life Sciences and a member of the research team. This is significant because by finding the metabolic pathway and understanding how the enzyme actually works and its structure gives us a blueprint of how allicin is created during biosynthesis.

Valentino and the team which included undergraduate students worked in the Sobrado Lab in the Fralin Life Sciences Institute directly with the substrates that comprise garlic, doing their work solely in vitro.

The researchers found that allicin, the component that gives garlic its smell and flavor, was produced by an entirely different biosynthetic process. Allyl-mercaptan reacts with flavin-containing monooxygenase, which then becomes allyl-sulfenic acid.

Importantly, the allicin levels can be tested, allowing farmers to know the strength of their crops without the need for genetic engineering. Greater flavor can simply be predicted, meaning powerful garlic could simply be bred or engineered.

We have a basic understanding of the biosynthesis of allicin that it is involved in flavor and smell, but we also now understand an enzyme that we can try to modulate, or a modify, to increase or decrease the level of the flavor molecules based on these biological processes, Sobrado said.

Because of their work, the future awaits for fields of garlic harsh enough to keep even the most terrifying vampires at bay.

There is a video with more information on this research.

Reference: Structure and function of a flavin-dependent S-monooxygenase from garlic (Allium sativum) by Hannah Valentino, Ashley C. Campbell, Jonathan P. Schuermann, Nazneen Sultana, Han G Nam, Sophie LeBlanc, John J. Tanner and Pablo Sobrado, 11 June 2020, Journal of Biological Chemistry.DOI: 10.1074/jbc.RA120.014484

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Discovery Could Lead to More Potent Garlic, Boosting Flavor and Aroma - SciTechDaily

COVID-19 was created in the Wuhan laboratory and is now in the hands of the Chinese military – Malaysian Christian News

COVID-19, which is killing and infecting people all over the world, is not a naturally occurring virus; instead, it was created in Wuhan, in a level-4 biosafety laboratory. Not only Chinese, but also French and US scientists contributed to the production of this "chimera," an organism created in a lab. Aug 08, 2020

By Bernardo CervelleraCOVID-19, which is killing and infecting people all over the world, is not a naturally occurring virus; instead, it was created in Wuhan, in a level-4 biosafety laboratory. Not only Chinese, but also French and US scientists contributed to the production of this "chimera," an organism created in a lab.

Until a few months ago, such an idea would have been labelled a conspiracy theory, contemptuously dismissed by people who believed in Chinas innocence, treated as absurd by several scholars who defended the "innocence" of science.

Now such thesis is presented with extensive documentation, dates, facts, names by an internationally renowned scientist, Prof Joesph Tritto, president of the Paris-based World Academy of Biomedical Sciences and Technologies (WABT), a non-governmental institution founded in 1997 under the auspices of UNESCO.

Prof Tritto, 68, is the author of Cina COVID 19. La Chimera che ha cambiato il Mondo (China COVID 19. The chimera that changed the world), a book that was released today by Edizioni Cantagalli.

In its 272 pages, which can be read like a thriller, Prof Tritto explains the origins of the virus with precision and scientific resolve, starting from the Chinese attempt to study vaccines against SARS, inserting genomes from HIV into organisms (which makes them more aggressive), adding elements of coronavirus discovered in horseshoe bats, using a method called reverse genetics system 2.

Prof Shi Zheng Li, head of the Wuhan laboratory, was the leading figure in these genetic engineering experiments, but the lab received help from the French government and the Pasteur institute, from which the Chinese learnt how to use HIV genomes.

Some US scientists also helped, including Prof Ralph S. Baric, of the University of North Carolina, with funds from the United States Agency for International Development (USAID). US scientists were interested in studying coronaviruses, which was prohibited in their country until 2017 due to their dangerousness.

Prof Tritto has a respectable curriculum. He is a doctor in urology, andrology, infertility microsurgery, and a professor in microtechnology and nanotechnology in the United Kingdom and India. He is a visiting professor and director of nano-medicine at Amity University in New Delhi (India).

For this very reason he could closely vet the research done in Wuhan. In his view, the latter began as a way to fight disease, but gradually turned into bioengineering studies to build lethal biological weapons.

It is no coincidence that in the past five years, the Wuhan lab received the largest portion of Chinas funding for virologic research, turning it into an advanced research centre under the direct control of the Chinese Academy of Sciences and the Chinese government.

According to Prof Tritto, Prof Shi Zheng-Li probably had no interest in doing work for the military or other purposes, unless she was forced to do so. Nobody doubts her good faith.

However, the fact remains that after the laboratory came into the spotlight because of the pandemic, Peoples Liberation Army Major General Chen Wei, an expert in biochemical weapons and bioterrorism, was appointed head of the Wuhan Institute of Virology, working with a team that includes Zhong Nanshan, a famous pulmonologist with long experience in infectious lung diseases.

In effect, the Wuhan Institute of Virology was placed under the control of Chinas military. Since then, nothing is known about Professor Shi Zheng-Li who seems to have disappeared.

In Trittos book, scientists come out badly. Driven by the desire for knowledge, they become eager for power, ambition, careerism and money.

Part of the book is devoted to vaccine research, in which institutes and labs compete against one another, not for the sake of medicine and to save the millions of coronavirus patients, but only to be the first to sell vaccines around the world.

China is well placed in this area. According to Prof Tritto, Beijing has only released partial data, and not made available the original genetic structure of the coronavirus (mother virus). Why? Because only with the original structure of the virus is it possible to produce a truly universal vaccine, effective everywhere on earth. Over time, viruses mutate and a vaccine produced by a mutated virus is effective only during a certain period time and in certain areas.

In other words, narrow commercial interests are the driving force, not love for science. Yet, lest we forget and Prof Tritto does not there are many heroes in this pandemic. In addition to the doctors and nurses who gave their lives to treat patients when they arrived in great numbers in emergency wards, we must cite the first doctors who reported the epidemic in Wuhan, who were forced into silence by police, threatened with dismissal.

One of these people is Dr Ai Fen, who reported a "strange influence" as early as November and was muzzled by hospital authorities. Another one is Dr Li Wenliang, an ophthalmologist who was forced to keep quiet and then died of COVID-19, infected by one of his patients. Nothing is currently known of Dr Ai Fen, who is missing.

Prof Trittos book also goes after the World Health Organisation, which has become - according to many - a "puppet" in Chinas hands, complicitous in its silence on the epidemic.

However, the book is not only turned to the past. Prof Tritto wants to see worldwide rules adopted for chimera research, level-4 biosafety labs, and cooperation between military and civil labs. China and other countries should also be forced to sign the Biological and Toxin Weapons Convention (BTWC).Asia News

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COVID-19 was created in the Wuhan laboratory and is now in the hands of the Chinese military - Malaysian Christian News

Training neural circuits early in development improves response, study finds – University of Illinois News

CHAMPAIGN, Ill. When it comes to training neural circuits for tissue engineering or biomedical applications, a new study suggests a key parameter: Train them young.

Techniques for training engineered neural circuits usually involve training them after the cells have fully matured. Using light-sensitive neurons derived from mouse stem cells, researchers at the University of Illinois, Urbana-Champaign found that training them throughout early cell development and network formation led to lasting improvements in the connections, responsivity and gene expression of the resulting neural network. They published their results in the journal Scientific Reports.

Its like an old dog learning new tricks versus a young puppy, said graduate student Gelson Pagan-Diaz, the first author of the study. When were training a network, if we stimulate it when its still like a puppy, we can get a better response to the training than if it were already mature.

Improved neural training has many applications in bioengineering and regenerative medicine. For example, the Illinois team hopes to use trained neural circuits to control the movement and behavior of miniature bio-hybrid machines. The types of improvements yielded by early training could give the machines and circuits more functionality and give the researchers more precise control over those functions.

As we advance the field of building machines with living cells, being able to stimulate and program neuronal cells and networks with light early in their development could be an important tool in our engineering repository, said study leader Rashid Bashir, a professor of bioengineering and dean of the Grainger College of Engineering at Illinois. Furthermore, this work could have implications for developmental biology, regenerative medicine and brain research.

To train the neurons, the researchers used timed pulses of light to stimulate the cells. The researchers began the training regimen when the cells were early in their development clusters of stem cells, called embryoid bodies, primed to become motor neurons. They continued the training as the cells differentiated, becoming fully mature neurons, and further continued it after transferring the cells to plates to connect and form neural circuits.

They then compared the early trained circuits with those cultured first and trained later the usual method.

The researchers saw a number of differences between the groups, Pagan-Diaz said. In the neurons trained during development, they saw more extensions indicating connections between cells, an increase in neurotransmitter packages sent between cells, and more structured nerve firing, indicating greater network stability. The effects of the early training were long-lasting, whereas cells trained later tended to have transient responses.

You can think of the neurons being like athletes in training, Pagan-Diaz said. The light stimulation was like a regular workout for the neurons they were stronger and more athletic, and did their jobs better.

To determine the underlying basis for these changes, the researchers analyzed the neurons genetic activity. They saw an increase in gene expression for genes related to network maturity and neural function, indicating that the early training could have permanently altered genetic pathways as the cells developed, Bashir said.

The researchers are continuing to explore what kinds of activities could be enhanced or programmed by early neuron training in the embryoid body phase. Embryoid bodies could be useful building-block components for biological machines, Pagan-Diaz said, and also hold promise for regenerative medicine.

Previous studies have shown that embryoid bodies with motor neurons implanted into mice that had been injured could improve the regeneration of tissue, Pagan-Diaz said. If we can improve or enhance the functionality of these embryoid bodies prior to putting them into an injured model, then theoretically we could enhance the recovery beyond what has been seen with injecting them and then stimulating them later.

The National Science Foundation supported this work through the Emergent Behaviors of Integrated Cellular Systems science and technology center and through the Miniature Brain Machinery Research Traineeship. Research staffer Jenny Drnevich, graduate students Karla Ramos-Cruz and Richard Sam, and University of Illinois, Chicago bioengineering professor Perijat Sengupta were co-authors of the paper.

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Training neural circuits early in development improves response, study finds - University of Illinois News

A Unicorn Rivalry Shaping the Future of the Synbio Industry – SynBioBeta

Two companies Ginkgo Bioworks and Zymergen are poised to shape the future of SynBio

Across industries as diverse as electronics, pharmaceuticals, food, and materials, industrial synthetic biology has begun to produce the goods we need more efficiently and sustainably. Using the synthetic biology approachor synbio for those familiar with the fieldstart-ups and Fortune 500 corporations alike are genetically-engineering microbes to manufacture a massive range of goods from the touchscreens in your phone, to the cotton in your t-shirt, to the meat in your plant-based burger.

Traditionally, this synthetic biology process has been a slow and hypothesis-driven endeavor, dependent on researcher geniusand a healthy dose of luckto design these microbial mini-factories. But not anymore.

Two companies have embraced the power of computational biology to take the guesswork out of synthetic biology and drive this young industry into the mainstream. Zymergen and Ginkgo Bioworks both use vast metagenomic databases, machine learning, and robust automated laboratories to design microbes custom-suited to manufacture a desired good. Both organizations have raised significant capital to build out their platforms, with Zymergen having raised a $400-million Series C in 2018 and Ginkgo Bioworks raising almost double that figure in multiple rounds over its lifetime.

Despite their shared mission to turn the art of engineering biology into a science, the two synbio pioneers are strongly differentiated by their respective technology stacks and business models. Analyzing the key differences between the two can help us understand how the synthetic biology industry as a whole may evolve and where the value levers are hidden in the synbio ecosystem.

At its core, industrial synthetic biology offers a path to both producing humanitys typical array of materialssuch as plastics, textiles, food, and medicinesmore efficiently, as well as creating completely new substances. To produce these goods with biology relies on a three-step process of:

1) Molecule Identification: Identifying the molecule you want to create;

2) Biology Design: Designing a microbe to manufacture the good; and

3) End-Product Manufacturing: Scaling the manufacturing.

Overlaying this simplified three-step value chain against each of these organizations core competencies illustrates where the two companies hope to compete in the industrial synbio field and how they will likely co-exist (or not) with other players in the space.

Both Zymergen and Ginkgo have established wide competitive moats around the Biology Design process. By constructing massive and growing metagenomic databasesmapping genes, proteins, and metabolic pathwaysand investing capital into automated labs capable of running high-throughput experimentation, the two companies have achieved a level of take-off velocity insulating them from would-be challengers seeking to enter the field.

Both companies have embraced an R&D process that creates a positive feedback loop, perpetually self-improving their ability to design microbes and putting greater distance between them and any potential competitors. Both companies leverage their massive datasets to train machine-learning algorithms that help design potential microbes for a given purpose. They then turn to their automated labs to test thousands of strains for performance, creating new data-points to further improve their machine-learning for the next batch of microbes.

While important differences exist between the two companies Biology Design stacks regarding the size of their datasets, the robustness of their software, and the automation of their hardware, both can claim strong proficiency in the Biology Design portion of the synbio value chain.

Beyond Biology Design, Ginkgo and Zymergen have taken divergent approaches to both Molecule Identification and End-Product Manufacturing. Regarding the former, Zymergen has developed chemistry and materials datasets and supporting lab infrastructure to rigorously scan their databases for promising molecules, predict their fit for a given purpose, and test those predictions empirically. To its credit, Zymergen has announced three products so fara biofilm for electronics, an insect repellent, and a crop pest control agentthat are all reported to be completely novel, never-used-before molecules.

Zymergen has also endeavored to internalize End-Product Manufacturing capabilities. After Zymergen designs a product, they turn to a network of close bio-manufacturing partners to produce the end-product in-house for downstream customers and partners.

While exceptions exist, Ginkgo has concentrated its focus almost solely on the Biology Design pieceperhaps with good reason. The ability to predict how a new molecule will behave varies across applications and necessitates a diverse toolkit: the analysis of a molecule to be used in electronic consumables varies greatly from that of one to be used in food or nutrition, diluting the scalability of a Molecule Identification skillset. For this reason, Ginkgo has focused their platform less on the discovery of totally novel substances, but more on finding a better means of manufacturing those we already know.

In regard to End-Product Manufacturing, commercial bio-manufacturing represents a fairly old technologyit relies primarily on the same fermentation process used to make beer.

Accordingly, Ginkgo believes that the End-Product Manufacturing element of the synbio process is relatively commoditized, and that their platform does not provide much marginal advantage here. In Ginkgos worldview, Molecule Identification appears to be too specialized beyond their core competency, while End-Product Manufacturing is perhaps too commoditized to offer much additional value.

In a nutshell, Zymergen has positioned itself as a products company with the infrastructure to cover the full product development process, while Ginkgo acts as a platform providing a modular solution at one valuable link in the synbio value chain. Under this strategy, Zymergen can produce differentiated products that cannot be found anywhere elseand Zymergen will be able to charge for that unique value.

Lacking an internal products engine, Ginkgo has cast a wide net of partnerships with industry innovators and internal spin-outs, in effect creating a decentralized product discovery network that can leverage its biology design platform to bring products to market.

While the two companies early strategies may not be indicative of their future evolution, their current directions do reflect their intended role within the larger synthetic biology ecosystem. On the one hand, Zymergen seems to be driving for a world where they can act as the one-stop-shop for anyones manufacturing or materials needsthey can ID the best substance, engineer the best organisms to produce it, and manufacture the product for sale to customers.

Alternatively, Ginkgos platform allows for a more distributed ecosystemwhere they can leverage their unique expertise in organism design to provide downstream partners with the crucial key to implementing their own bio-manufacturing processes.

If either approach can succeed in pulling synthetic biology into the mainstream, we could all benefit from a cleaner, healthier, and more abundant world.

Matthew Kirshner works in life sciences consulting at Putnam Associates, focused on pharma and biotech organizations commercializing novel therapeutics and diagnostics. He has no professional affiliation in his work with either company referenced in this article.

Originally published on Genetic Engineering & Biotechnology News https://www.genengnews.com/gen-edge/a-unicorn-rivalry-shaping-the-future-of-the-synbio-industry/

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A Unicorn Rivalry Shaping the Future of the Synbio Industry - SynBioBeta

Protests have erupted across the nation and returned to St. Louis: Here are some of the leaders of the movement – STLtoday.com

The Rev. Darryl Gray sits at the intersection of Market Street and Tucker Boulevard for minutes of silence in memory of those killed by police officers, near City Hall in downtown St. Louis on Wednesday, July 8, 2020. Protesters announced plans to camp in front of the building until their demands were met and Mayor Lyda Krewson resigns. Photo by Robert Cohen, rcohen@post-dispatch.com

Protests dont just happen as spontaneous moments.

They are planned and executed by an array of people college students and politicians, lawyers and social workers, medical professionals and blue-collar workers spurred by outrage, activism or a sense of injustice.

Protesters who emerged as leaders six years ago after 18-year-old Michael Brown was shot and killed by a Ferguson police officer have in some cases carried their commitment in new directions of influence, including freshman state Rep. Rasheen Aldridge and Cori Bush, the Democratic nominee in the 1st Congressional District after Tuesdays primary election.

Aldridge and Bush have remained active in protests, including those sparked this summer by the death of George Floyd, a Black man killed while restrained by a Minneapolis police officer, as well as by the release of a video that showed a Florissant police detective hitting a man with his vehicle.

But they are not the only leaders of the movement.

At the center of the demonstrations are a mix of groups ExpectUs, RespectUs, Tent Mission STL, Occupy City Hall STL, Protest THAT, Action St. Louis and ClosetheWorkhouse whose opinions vary on the best way to seek justice.

And on the anniversary of Browns death, Aug. 9, 2014, protest leaders say underlying inequities and a lack of broad reform measures make the St. Louis area ripe for social unrest.

Meet a few of the leaders who carry the culture of protest in the St. Louis region.

The pastor

The Rev. Daryl Gray, 66, is a founder of the protest group ExpectUs. He is one of several Democratic leaders in the organization who doubles as both an activist and a politician. Others include Aldridge and Bush, who toppled longtime U.S. Rep. William Lacy Clay in Tuesdays Democratic primary.

Among other calls for justice, ExpectUs has advocated for reparations for Black people and an end to police brutality. Gray said current protests are more of a movement than a moment.

Rev. Darryl Gray talks to police officers as protesters march at the Galleria in Richmond Heights on Black Friday, Nov. 24, 2017. About 100 people marched through the mall, calling for an economic boycott of area businesses. Photo by Robert Cohen, rcohen@post-dispatch.com

Born in Boston and having grown up in both South Carolina and Canada, Gray moved to St. Louis after Fergusons uprisings. He emphasizes that when an organization like ExpectUs is grassroots, its bound to have more buy-in from its members.

If you own it, then youre going to go the extra mile for it, Gray said.

Gray attended a newly integrated high school in South Carolina, where he got a taste of standing up for what he believed in and the repercussions that often follow.

During a school pep rally, Gray said a man came riding into the gym on a horse waving a Confederate flag. He walked out, and hundreds of students followed him. He was suspended from school, he said, for inciting a disturbance.

I didnt know they were behind me. I didnt look back, he said.

Gray went on to join the U.S. Army, where he was honorably discharged. He then campaigned for presidential candidate the Rev. Jesse Jackson in Florida, worked with the Southern Christian Leadership Conference, preached in both Canada and Atlanta, served as head of Atlantas NAACP, and briefly served as a state senator from Kansas City.

Now, he said, hes ready to make St. Louis his final destination.

With a history of frequently moving, proving himself in new cities has been the norm for Gray.

I think Ive gotten beat up enough in the streets. Ive shown up more than some people whove been here all their lives thats got to count for something, Gray said.

On July 6, a day after protesters say they were beaten while being arrested for protesting at Florissants police department, Gray asked officers in riot gear behind the gate of the police department to go back inside the police department as a sign of good faith. A white shirt officer agreed and instructed riot police to go inside, but a regular, uniformed group of police then came out another door.

Gray and other ExpectUs leaders told everyone to go home. Some in the crowd strongly opposed leaving though, and the two factions argued with one another.

That was our mistake the timing. For us to disperse while police were standing there could be perceived as a sign of weakness, Gray said. RespectUs felt disrespected at that moment, and they had reason to. Our timing was off. That was our mistake. We owned it.

The self-proclaimed radical

One of the protesters who stayed behind that night was Tauren Taylor, a 25-year-old University City resident.

A self-proclaimed radical, Taylor said he stayed because protesters were supposed to go out on their own terms. Taylor said protesters had to advocate for every inch of real estate on which to protest, including the parking lot across the street from the police station.

Tauren Taylor, center, talks with Dhoruba Shakur, left, and Tory Russell as they block the doorway to the Ferguson Market to shut it down for the night on Thursday, July 30, 2020, after St. Louis County prosecutor Wesley Bell announced that after a review he would not charge police officer Darren Wilson in Brown's 2014 death. The store became a central part of the case after the teenager was accused of robbing it before he was shot. Photo by Robert Cohen, rcohen@post-dispatch.com

Why keep letting them hand us stuff when our job is to take things? Were supposed to be forcing them to give us things they dont want to give us, Taylor said. They dont want to give us freedom. They dont want to stop killing us, they dont want to stop beating us.

Taylor has been arrested at several protests in the past year. He was also one of the protest leaders after Terry Tillman, 23, was killed last year by police near the St. Louis Galleria Mall in Richmond Heights.

Taylor remains vocal on the front line, but he says he will not join a protest group.

To be held under a name, youre held under their standards. ExpectUs, for instance, I love people in ExpectUS. I will ride for ExpectUs, but I dont think now is the time for us to try to ask for reparations.

Taylor moved to Missouri from California when he was a teenager. He said his family dealt with poverty, and he was bullied frequently for having a lisp and being studious.

One of his first successful protests, he said, was getting a neglectful teacher fired by collecting petition names.

Once I found out what the chain of the command was, I went up that ladder, he said.

Eventually, he said, he was kicked out of Vashon High School for fighting.

Taylor earned a diploma from the Fresh Start Academy program at 17, and briefly studied animal science and genetic engineering at St. Louis Community College at Forest Park.

He acknowledged some may see his protest methods as extreme, but he said his methods also help people understand someone is sticking up for them.

Taylor said hes not sure about a future career path, but hes working odd jobs, including selling homemade goods at Soulard Market. In the meantime, he said, he will continue to protest injustices wherever they spring up in the St. Louis area.

Tauren Taylor leads chants in the parking lot of the Florissant Police Department on Monday, July 6, 2020. Protesters held a demonstration against police brutality after a Florissant officer was fired and charged with hitting a fleeing subject with his vehicle. Those attending were seeking charges against two other officers involved in the incident. Photo by Robert Cohen, rcohen@post-dispatch.com

Taylor was one of several protesters to take exception with comments in mid-July by Jimmie Edwards, the top law enforcement official in St. Louis. Edwards, at an anti-crime demonstration outside City Hall, said many protesters werent from the city, and he condemned violence that erupted at some of the protests.

I feel like violence happens everywhere, even in nature, said Taylor. Were all human, we all make mistakes. Theres no way to control every single person thats out there. Ive seen police hurt people and each other. You cant condone it on one end and condemn it on the other.

The elder

The perceived lack of government response when doorbell security footage showed former Florissant Detective Joshua Smith running over a fleeing suspect in early June led to the formation of another protest group, RespectUs, said one of the founders, Cathy Mama Cat Daniels.

Daniels, 59, said she and a group of other front-line Ferguson protesters went to the police station for answers about what had happened, but there were barricades around the police department.

Protester Cathy Daniels leads the chant opposing the King Louis IX statue in Forest Park, during a rally atop Art Hill on Saturday, June 27, 2020. About 200 people on opposite sides of the debate attended the rally in front of the St. Louis Art Museum. Photo by Robert Cohen, rcohen@post-dispatch.com

When they didnt answer us and treat us with the human respect as people who live in this town, youve got to stand up and fight back, said Daniels. Even the (city) council, anyone who spoke against their idea of democracy, they didnt feel the need to respect them. Thats why we are RespectUs.

Daniels said RespectUs has four core demands: fire, charge, arrest and convict Smith. Thus far, three of those four have happened Smith has not yet had his day in court.

Daniels has lived in Florissant since 2012. She grew up in New York City and previously lived in Chicago and San Diego. Since Fergusons uprisings, shes worked as a cook and founded PotBangerz, a nonprofit dedicated to providing food and clothing to families in need. The organization is now renovating a home in Pine Lawn for cis, queer and trans women in need and plans to open the home later this year.

Cathy 'Mama Cat' Daniels empties one of two massive bowls of salad with vegetables donated from area farms, as she cooks dinner for 130 people in the kitchen at First Congregational Church of St. Louis on Thursday, July 23, 2020 in Clayton. Daniels leads the PotBangerz, a group of volunteers, many who are active in area protests, who feed the hungry and the unhoused. Photo by Robert Cohen, rcohen@post-dispatch.com

An elder of the protest group, Daniels said she offers advice to protesters and watches from the back, cane and chair in tow. Still, she said, she doesnt always approve of the protest methods that unfold.

Theres no such thing as peaceful protest. Thats an oxymoron. If there was peace, we wouldnt need to protest, Daniels said. I dont support tearing up stuff. We had that happen during Ferguson. If were gonna burn it down, then do it the right way, and thats not in the literal sense. Burn down the system. Defund the police.

The congresswoman

Daniels stood behind Cori Bush, 44, as hundreds of protesters returned July 3 to Portland Place in St. Louis West End. The private street had recently made national news, as Mark and Patricia McCloskey waved guns at protesters in an effort to, as they said, defend their home. The couple has since been charged with unlawful use of a weapon.

There was nothing to defend, and thats what angers me so bad, Bush said.

Protest leader Cori Bush marches on Forest Park Parkway during a rally and march against police brutality in the Delmar Loop on Friday, June 12, 2020. Photo by Robert Cohen, rcohen@post-dispatch.com

In addition to rising as a leader of ExpectUs, Bush gained national notoriety in Tuesdays election. In a district that has historically voted overwhelmingly for Democrats, Bush will likely become the first Black woman to represent Missouri in Congress.

Bush said she learned about politics, protest tactics and demonstrations from her father, Earl Bush, who worked as a politician in north St. Louis County.

She attended high school at Cardinal Ritter College Prep and said she had no intention of starting a life in politics. She graduated from Harris- Stowe State University and Lutheran School of Nursing, then entered the fields of nursing and ministry.

Between Bushs unsuccessful runs for Senate and the House of Representatives in 2016 and 2018, respectively, St. Louis police officer Jason Stockley went on trial for shooting a Black man, Anthony Lamar Smith, in 2011. Stockley was charged with first-degree murder, and prosecutors claimed he planted a gun on Smith. When Stockley was found not guilty by a judge in 2017, protests erupted again, and Bush emerged as a leader of ExpectUs.

During those demonstrations, Bush said, the group began to change how it disseminated information.

With Ferguson, you didnt have to call and ask around to see what time a protest was going on. You could just show up. It was 24/7. With Stockley, we had to have a way to get the information out, said Bush, highlighting the organizations use of social media.

Now, both she and Gray agree the groups protests are more refined, pointing out the local history lessons they provide before the marching and chanting begin.

ExpectUs doesnt do violent things, but what were not going to do is turn our backs on people that do, as far as their form of protest, Bush said, referencing the violence that occasionally breaks out at protests, including an early June demonstration that included gunfire and widespread looting as the night wore on.

If I have a sandwich today for breakfast, lunch and dinner Im not going to loot a sandwich, Bush said. If they fix the problems, they wont have to worry about that.

Bush said shes aware she could get blamed for the violence that occurs during protests, but she stands by her decision to continually show up and call for change.

I know it every single time, and I make that choice to show up, she said. When we stop showing up, when we stop pushing, thats how they win, so Ill take that chance.

This story has been updated to correct the year of the shooting of Anthony Lamar Smith.

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Protests have erupted across the nation and returned to St. Louis: Here are some of the leaders of the movement - STLtoday.com

Discovery Could Lead to More Potent Garlic, Boosting Flavor and Aroma – SciTechDaily

Hannah Valentino, left, and Pablo Sobrado, right, uncover a new step in the process that makes garlic potent. Credit: Virginia Tech

For centuries, people around the world have used garlic as a spice, natural remedy, and pest deterrent but they didnt know how powerful or pungent the heads of garlic were until they tasted them.

This information changes the whole story about how garlic could be improved. Hannah Valentino, a Ph.D. candidate in the Virginia Tech College of Agriculture and Life Sciences

But what if farmers were able to grow garlic and know exactly how potent it would be? What if buyers could pick their garlic based on its might?

A team of Virginia Tech researchers recently discovered a new step in the metabolic process that produces the enzyme allicin, which leads to garlics delectable flavor and aroma, a finding that upends decades of previous scientific belief. Their work could boost the malodorous yet delicious characteristics that garlic-lovers the world over savor.

This information changes the whole story about how garlic could be improved or we could make the compounds responsible for its unique flavor, said Hannah Valentino, a College of Agriculture and Life Sciences Ph.D. candidate. This could lead to a new strain of garlic that would produce more flavor.

The discovery of this pathway opens the door for better control of production and more consistent crops, which would help farmers. Garlic could be sold as strong or weak, depending on consumer preferences.

The research was recently published in the Journal of Biological Chemistry.

Hannah Valentino, left, and Pablo Sobrado, right, are conducting research that is laying the foundation for a future in which buyers can choose garlic based on its strength and flavor profile. Credit: Virginia Tech

When Valentino, an Institute for Critical Technology and Applied Science doctoral fellow, and her team set out to test the generally accepted biological process that creates allicin, they found it just didnt happen.

Thats when the team of researchers set out to discover what was really happening in garlic.

As they peeled back the layers, they realized there was no fuel to power the previously accepted biological process that creates allicin.

By using rational design, Hannah found a potential substrate, said Pablo Sobrado, professor of biochemistry in the College of Agriculture and Life Sciences and a member of the research team. This is significant because by finding the metabolic pathway and understanding how the enzyme actually works and its structure gives us a blueprint of how allicin is created during biosynthesis.

Valentino and the team which included undergraduate students worked in the Sobrado Lab in the Fralin Life Sciences Institute directly with the substrates that comprise garlic, doing their work solely in vitro.

The researchers found that allicin, the component that gives garlic its smell and flavor, was produced by an entirely different biosynthetic process. Allyl-mercaptan reacts with flavin-containing monooxygenase, which then becomes allyl-sulfenic acid.

Importantly, the allicin levels can be tested, allowing farmers to know the strength of their crops without the need for genetic engineering. Greater flavor can simply be predicted, meaning powerful garlic could simply be bred or engineered.

We have a basic understanding of the biosynthesis of allicin that it is involved in flavor and smell, but we also now understand an enzyme that we can try to modulate, or a modify, to increase or decrease the level of the flavor molecules based on these biological processes, Sobrado said.

Because of their work, the future awaits for fields of garlic harsh enough to keep even the most terrifying vampires at bay.

There is a video with more information on this research.

Reference: Structure and function of a flavin-dependent S-monooxygenase from garlic (Allium sativum) by Hannah Valentino, Ashley C. Campbell, Jonathan P. Schuermann, Nazneen Sultana, Han G Nam, Sophie LeBlanc, John J. Tanner and Pablo Sobrado, 11 June 2020, Journal of Biological Chemistry.DOI: 10.1074/jbc.RA120.014484

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Discovery Could Lead to More Potent Garlic, Boosting Flavor and Aroma - SciTechDaily

Training neural circuits early in development improves response, study finds – University of Illinois News

CHAMPAIGN, Ill. When it comes to training neural circuits for tissue engineering or biomedical applications, a new study suggests a key parameter: Train them young.

Techniques for training engineered neural circuits usually involve training them after the cells have fully matured. Using light-sensitive neurons derived from mouse stem cells, researchers at the University of Illinois, Urbana-Champaign found that training them throughout early cell development and network formation led to lasting improvements in the connections, responsivity and gene expression of the resulting neural network. They published their results in the journal Scientific Reports.

Its like an old dog learning new tricks versus a young puppy, said graduate student Gelson Pagan-Diaz, the first author of the study. When were training a network, if we stimulate it when its still like a puppy, we can get a better response to the training than if it were already mature.

Improved neural training has many applications in bioengineering and regenerative medicine. For example, the Illinois team hopes to use trained neural circuits to control the movement and behavior of miniature bio-hybrid machines. The types of improvements yielded by early training could give the machines and circuits more functionality and give the researchers more precise control over those functions.

As we advance the field of building machines with living cells, being able to stimulate and program neuronal cells and networks with light early in their development could be an important tool in our engineering repository, said study leader Rashid Bashir, a professor of bioengineering and dean of the Grainger College of Engineering at Illinois. Furthermore, this work could have implications for developmental biology, regenerative medicine and brain research.

To train the neurons, the researchers used timed pulses of light to stimulate the cells. The researchers began the training regimen when the cells were early in their development clusters of stem cells, called embryoid bodies, primed to become motor neurons. They continued the training as the cells differentiated, becoming fully mature neurons, and further continued it after transferring the cells to plates to connect and form neural circuits.

They then compared the early trained circuits with those cultured first and trained later the usual method.

The researchers saw a number of differences between the groups, Pagan-Diaz said. In the neurons trained during development, they saw more extensions indicating connections between cells, an increase in neurotransmitter packages sent between cells, and more structured nerve firing, indicating greater network stability. The effects of the early training were long-lasting, whereas cells trained later tended to have transient responses.

You can think of the neurons being like athletes in training, Pagan-Diaz said. The light stimulation was like a regular workout for the neurons they were stronger and more athletic, and did their jobs better.

To determine the underlying basis for these changes, the researchers analyzed the neurons genetic activity. They saw an increase in gene expression for genes related to network maturity and neural function, indicating that the early training could have permanently altered genetic pathways as the cells developed, Bashir said.

The researchers are continuing to explore what kinds of activities could be enhanced or programmed by early neuron training in the embryoid body phase. Embryoid bodies could be useful building-block components for biological machines, Pagan-Diaz said, and also hold promise for regenerative medicine.

Previous studies have shown that embryoid bodies with motor neurons implanted into mice that had been injured could improve the regeneration of tissue, Pagan-Diaz said. If we can improve or enhance the functionality of these embryoid bodies prior to putting them into an injured model, then theoretically we could enhance the recovery beyond what has been seen with injecting them and then stimulating them later.

The National Science Foundation supported this work through the Emergent Behaviors of Integrated Cellular Systems science and technology center and through the Miniature Brain Machinery Research Traineeship. Research staffer Jenny Drnevich, graduate students Karla Ramos-Cruz and Richard Sam, and University of Illinois, Chicago bioengineering professor Perijat Sengupta were co-authors of the paper.

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Training neural circuits early in development improves response, study finds - University of Illinois News

COVID-19 was created in the Wuhan laboratory and is now in the hands of the Chinese military – Malaysian Christian News

COVID-19, which is killing and infecting people all over the world, is not a naturally occurring virus; instead, it was created in Wuhan, in a level-4 biosafety laboratory. Not only Chinese, but also French and US scientists contributed to the production of this "chimera," an organism created in a lab. Aug 08, 2020

By Bernardo CervelleraCOVID-19, which is killing and infecting people all over the world, is not a naturally occurring virus; instead, it was created in Wuhan, in a level-4 biosafety laboratory. Not only Chinese, but also French and US scientists contributed to the production of this "chimera," an organism created in a lab.

Until a few months ago, such an idea would have been labelled a conspiracy theory, contemptuously dismissed by people who believed in Chinas innocence, treated as absurd by several scholars who defended the "innocence" of science.

Now such thesis is presented with extensive documentation, dates, facts, names by an internationally renowned scientist, Prof Joesph Tritto, president of the Paris-based World Academy of Biomedical Sciences and Technologies (WABT), a non-governmental institution founded in 1997 under the auspices of UNESCO.

Prof Tritto, 68, is the author of Cina COVID 19. La Chimera che ha cambiato il Mondo (China COVID 19. The chimera that changed the world), a book that was released today by Edizioni Cantagalli.

In its 272 pages, which can be read like a thriller, Prof Tritto explains the origins of the virus with precision and scientific resolve, starting from the Chinese attempt to study vaccines against SARS, inserting genomes from HIV into organisms (which makes them more aggressive), adding elements of coronavirus discovered in horseshoe bats, using a method called reverse genetics system 2.

Prof Shi Zheng Li, head of the Wuhan laboratory, was the leading figure in these genetic engineering experiments, but the lab received help from the French government and the Pasteur institute, from which the Chinese learnt how to use HIV genomes.

Some US scientists also helped, including Prof Ralph S. Baric, of the University of North Carolina, with funds from the United States Agency for International Development (USAID). US scientists were interested in studying coronaviruses, which was prohibited in their country until 2017 due to their dangerousness.

Prof Tritto has a respectable curriculum. He is a doctor in urology, andrology, infertility microsurgery, and a professor in microtechnology and nanotechnology in the United Kingdom and India. He is a visiting professor and director of nano-medicine at Amity University in New Delhi (India).

For this very reason he could closely vet the research done in Wuhan. In his view, the latter began as a way to fight disease, but gradually turned into bioengineering studies to build lethal biological weapons.

It is no coincidence that in the past five years, the Wuhan lab received the largest portion of Chinas funding for virologic research, turning it into an advanced research centre under the direct control of the Chinese Academy of Sciences and the Chinese government.

According to Prof Tritto, Prof Shi Zheng-Li probably had no interest in doing work for the military or other purposes, unless she was forced to do so. Nobody doubts her good faith.

However, the fact remains that after the laboratory came into the spotlight because of the pandemic, Peoples Liberation Army Major General Chen Wei, an expert in biochemical weapons and bioterrorism, was appointed head of the Wuhan Institute of Virology, working with a team that includes Zhong Nanshan, a famous pulmonologist with long experience in infectious lung diseases.

In effect, the Wuhan Institute of Virology was placed under the control of Chinas military. Since then, nothing is known about Professor Shi Zheng-Li who seems to have disappeared.

In Trittos book, scientists come out badly. Driven by the desire for knowledge, they become eager for power, ambition, careerism and money.

Part of the book is devoted to vaccine research, in which institutes and labs compete against one another, not for the sake of medicine and to save the millions of coronavirus patients, but only to be the first to sell vaccines around the world.

China is well placed in this area. According to Prof Tritto, Beijing has only released partial data, and not made available the original genetic structure of the coronavirus (mother virus). Why? Because only with the original structure of the virus is it possible to produce a truly universal vaccine, effective everywhere on earth. Over time, viruses mutate and a vaccine produced by a mutated virus is effective only during a certain period time and in certain areas.

In other words, narrow commercial interests are the driving force, not love for science. Yet, lest we forget and Prof Tritto does not there are many heroes in this pandemic. In addition to the doctors and nurses who gave their lives to treat patients when they arrived in great numbers in emergency wards, we must cite the first doctors who reported the epidemic in Wuhan, who were forced into silence by police, threatened with dismissal.

One of these people is Dr Ai Fen, who reported a "strange influence" as early as November and was muzzled by hospital authorities. Another one is Dr Li Wenliang, an ophthalmologist who was forced to keep quiet and then died of COVID-19, infected by one of his patients. Nothing is currently known of Dr Ai Fen, who is missing.

Prof Trittos book also goes after the World Health Organisation, which has become - according to many - a "puppet" in Chinas hands, complicitous in its silence on the epidemic.

However, the book is not only turned to the past. Prof Tritto wants to see worldwide rules adopted for chimera research, level-4 biosafety labs, and cooperation between military and civil labs. China and other countries should also be forced to sign the Biological and Toxin Weapons Convention (BTWC).Asia News

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COVID-19 was created in the Wuhan laboratory and is now in the hands of the Chinese military - Malaysian Christian News

A Unicorn Rivalry Shaping the Future of the Synbio Industry – SynBioBeta

Two companies Ginkgo Bioworks and Zymergen are poised to shape the future of SynBio

Across industries as diverse as electronics, pharmaceuticals, food, and materials, industrial synthetic biology has begun to produce the goods we need more efficiently and sustainably. Using the synthetic biology approachor synbio for those familiar with the fieldstart-ups and Fortune 500 corporations alike are genetically-engineering microbes to manufacture a massive range of goods from the touchscreens in your phone, to the cotton in your t-shirt, to the meat in your plant-based burger.

Traditionally, this synthetic biology process has been a slow and hypothesis-driven endeavor, dependent on researcher geniusand a healthy dose of luckto design these microbial mini-factories. But not anymore.

Two companies have embraced the power of computational biology to take the guesswork out of synthetic biology and drive this young industry into the mainstream. Zymergen and Ginkgo Bioworks both use vast metagenomic databases, machine learning, and robust automated laboratories to design microbes custom-suited to manufacture a desired good. Both organizations have raised significant capital to build out their platforms, with Zymergen having raised a $400-million Series C in 2018 and Ginkgo Bioworks raising almost double that figure in multiple rounds over its lifetime.

Despite their shared mission to turn the art of engineering biology into a science, the two synbio pioneers are strongly differentiated by their respective technology stacks and business models. Analyzing the key differences between the two can help us understand how the synthetic biology industry as a whole may evolve and where the value levers are hidden in the synbio ecosystem.

At its core, industrial synthetic biology offers a path to both producing humanitys typical array of materialssuch as plastics, textiles, food, and medicinesmore efficiently, as well as creating completely new substances. To produce these goods with biology relies on a three-step process of:

1) Molecule Identification: Identifying the molecule you want to create;

2) Biology Design: Designing a microbe to manufacture the good; and

3) End-Product Manufacturing: Scaling the manufacturing.

Overlaying this simplified three-step value chain against each of these organizations core competencies illustrates where the two companies hope to compete in the industrial synbio field and how they will likely co-exist (or not) with other players in the space.

Both Zymergen and Ginkgo have established wide competitive moats around the Biology Design process. By constructing massive and growing metagenomic databasesmapping genes, proteins, and metabolic pathwaysand investing capital into automated labs capable of running high-throughput experimentation, the two companies have achieved a level of take-off velocity insulating them from would-be challengers seeking to enter the field.

Both companies have embraced an R&D process that creates a positive feedback loop, perpetually self-improving their ability to design microbes and putting greater distance between them and any potential competitors. Both companies leverage their massive datasets to train machine-learning algorithms that help design potential microbes for a given purpose. They then turn to their automated labs to test thousands of strains for performance, creating new data-points to further improve their machine-learning for the next batch of microbes.

While important differences exist between the two companies Biology Design stacks regarding the size of their datasets, the robustness of their software, and the automation of their hardware, both can claim strong proficiency in the Biology Design portion of the synbio value chain.

Beyond Biology Design, Ginkgo and Zymergen have taken divergent approaches to both Molecule Identification and End-Product Manufacturing. Regarding the former, Zymergen has developed chemistry and materials datasets and supporting lab infrastructure to rigorously scan their databases for promising molecules, predict their fit for a given purpose, and test those predictions empirically. To its credit, Zymergen has announced three products so fara biofilm for electronics, an insect repellent, and a crop pest control agentthat are all reported to be completely novel, never-used-before molecules.

Zymergen has also endeavored to internalize End-Product Manufacturing capabilities. After Zymergen designs a product, they turn to a network of close bio-manufacturing partners to produce the end-product in-house for downstream customers and partners.

While exceptions exist, Ginkgo has concentrated its focus almost solely on the Biology Design pieceperhaps with good reason. The ability to predict how a new molecule will behave varies across applications and necessitates a diverse toolkit: the analysis of a molecule to be used in electronic consumables varies greatly from that of one to be used in food or nutrition, diluting the scalability of a Molecule Identification skillset. For this reason, Ginkgo has focused their platform less on the discovery of totally novel substances, but more on finding a better means of manufacturing those we already know.

In regard to End-Product Manufacturing, commercial bio-manufacturing represents a fairly old technologyit relies primarily on the same fermentation process used to make beer.

Accordingly, Ginkgo believes that the End-Product Manufacturing element of the synbio process is relatively commoditized, and that their platform does not provide much marginal advantage here. In Ginkgos worldview, Molecule Identification appears to be too specialized beyond their core competency, while End-Product Manufacturing is perhaps too commoditized to offer much additional value.

In a nutshell, Zymergen has positioned itself as a products company with the infrastructure to cover the full product development process, while Ginkgo acts as a platform providing a modular solution at one valuable link in the synbio value chain. Under this strategy, Zymergen can produce differentiated products that cannot be found anywhere elseand Zymergen will be able to charge for that unique value.

Lacking an internal products engine, Ginkgo has cast a wide net of partnerships with industry innovators and internal spin-outs, in effect creating a decentralized product discovery network that can leverage its biology design platform to bring products to market.

While the two companies early strategies may not be indicative of their future evolution, their current directions do reflect their intended role within the larger synthetic biology ecosystem. On the one hand, Zymergen seems to be driving for a world where they can act as the one-stop-shop for anyones manufacturing or materials needsthey can ID the best substance, engineer the best organisms to produce it, and manufacture the product for sale to customers.

Alternatively, Ginkgos platform allows for a more distributed ecosystemwhere they can leverage their unique expertise in organism design to provide downstream partners with the crucial key to implementing their own bio-manufacturing processes.

If either approach can succeed in pulling synthetic biology into the mainstream, we could all benefit from a cleaner, healthier, and more abundant world.

Matthew Kirshner works in life sciences consulting at Putnam Associates, focused on pharma and biotech organizations commercializing novel therapeutics and diagnostics. He has no professional affiliation in his work with either company referenced in this article.

Originally published on Genetic Engineering & Biotechnology News https://www.genengnews.com/gen-edge/a-unicorn-rivalry-shaping-the-future-of-the-synbio-industry/

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A Unicorn Rivalry Shaping the Future of the Synbio Industry - SynBioBeta

A versatile genetic control system in mammalian cells and mice responsive to clinically licensed sodium ferulate – Science Advances

Abstract

Dynamically adjustable gene- and cell-based therapies are recognized as next-generation medicine. However, the translation of precision therapies into clinics is limited by lack of specific switches controlled by inducers that are safe and ready for clinical use. Ferulic acid (FA) is a phytochemical with a wide range of therapeutic effects, and its salt sodium ferulate (SF) is used as an antithrombotic drug in clinics. Here, we describe an FA/SF-adjustable transcriptional switch controlled by the clinically licensed drug SF. We demonstrated that SF-responsive switches can be engineered to control CRISPR-Cas9 systems for on-command genome/epigenome engineering. In addition, we integrated FA-controlled switches into programmable biocomputers to process logic operations. We further demonstrated the dose-dependent SF-inducible transgene expression in mice by oral administration of SF tablets. Engineered switches responsive to small-molecule clinically licensed drugs to achieve adjustable transgene expression profiles provide new opportunities for dynamic interventions in gene- and cell-based precision medicine.

Gene- and cell-based therapies are rapidly emerging innovation drivers in modern medical sciences (1, 2). Mammalian synthetic biology is boosting this innovation by providing the necessary molecular tools and design strategies for cell-based precision medicine (3). A particular focus in cell-based precision therapy is on externally programmable gene switches to align target gene activities with complex therapeutic requirements in living cells and organisms (4). This is of particular interest with regard to improving therapeutic outcome while reducing possible side effects. For example, there is a high demand for on-command switchable CRISPR (clustered regularly interspaced short palindromic repeats)Cas9 systems to achieve desired genome/epigenome editing while avoiding off-target effects due to prolonged, unspecific Cas9 activity (5, 6). Furthermore, despite the observation that genetic engineered chimeric antigen receptor T (CAR-T) cells are achieving unprecedented response rates against leukemia (7), there is the urgent need to improve treatment safety and reliability by integrating sophisticated control switches to avoid side effects such as the cytokine release syndrome (8). Obviously, the further advancement of such promising therapeutic technologies is currently severely limited by a lack of safe and specific control systems to tune transgene activity and regulate cell behavior in a dynamic, patient-tailored fashion.

Initial efforts in the design of dynamically controlled genetic switches relied on antibiotics [e.g., tetracycline (9), streptogramins (10), or macrolides (11)] as triggers to control cell behaviors. Later on, genetic switches were engineered to be activated by cosmetic compounds [e.g., phloretin (12) and paraben (13)], food additives [e.g., vanillic acid (14) and benzoate (BA) (15)], and even food components [e.g., caffeine (16), spearmint (17), and protocatechuic acid (18)]. However, these inducers either can lead to antibiotic resistance, are not yet clinically approved for long-term use in patients, or are present in common food leading to undesired switch activation. Physical inducers, such as light (19, 20), magnetic force (21), and temperature (22), have recently been developed to control the expression and release of therapeutic outputs. However, specialized hardware and software are often required for administration of these stimuli. Furthermore, fluctuations of the physical inducers in everyday life make them less orthogonal and likely lead to unwanted interference. For these reasons, ideal inducers would act in a specific manner, would have no toxic effects, would be safe and clinically approved for human use, and would be easily obtained from natural sources (23). Genetic switches responsive to such ideal inducers would show substantially increased chances for translation into clinical practice (24).

Ferulic acid (FA) is a widely distributed hydroxycinnamic acid, an abundant polyphenolic compound found in various vegetables, fruits, and grain, exhibiting potent antioxidant and various therapeutic activities (25). Because of its strong antioxidant activity by scavenging free radicals and activating antioxidant enzymes, it has been widely applied to prevent reactive oxygen speciesrelated diseases (26), such as cancer (27), cardiovascular diseases (28), and diabetes mellitus (29). It is also approved as a food additive in some countries (30). Its sodium salt, sodium ferulate (SF), is a compound widely used in traditional Chinese medicine for several decades and is approved by the National Medical Products Administration (NMPA) for treating cardiovascular and cerebrovascular diseases and to prevent thrombosis (3133). Overall, FA/SF has all it takes to be a safe and physiologically inert inducer for genetic switches designed for future therapeutic applications.

It has been reported that FA can trigger dissociation of the phenolic acid decarboxylase regulator (PadR) repressor, a transcriptional regulator of the padA gene in Bacillus subtilis 168, from its specific DNA binding sequence OPadR (34, 35). On the basis of this switchable molecular interaction, we propose to construct a novel genetic switch in which PadR is fused to eukaryotic epigenetic effector domains. This synthetic transcription factor would specifically modulate the activity of synthetic promoters containing the PadR-specific OPadR operator in response to varying concentrations of FA/SF. We here present the FA/SF-controlled, orthogonal genetic switches termed FAR (FA regulator) that are responsive to the clinical drug SF to fine-tune transgene expression in various mammalian cell lines and in mice. We further demonstrate applicability of the designed FAR switches for multiple applications: (i) in combination with CRISPR-Cas9 for inducible gene editing and epigenome engineering, (ii) as modular building block for engineering programmable biocomputation to perform five logic operations in single mammalian cells, and (iii) the ability to program transgene expression in mice in response to oral administration of clinically approved SF tablets. This versatility combined with robust switch characteristics make the FAR switches a potent instrument for cybergenetics and expand the molecular intervention strategies that may foster novel advances in gene- and cell-based precision therapies.

To engineer a mammalian cellcompatible FA-controlled OFF switch, where gene expression is switched off in the presence of FA, we constructed a synthetic mammalian transcription activator, the activation version of phenolic acid decarboxylase regulator (aPadR), by fusing a VP64 transactivation domain (four copies of the Herpes simplex virus protein 16) to the N terminus of PadR (VP64-PadR). The transactivator aPadR binds and initiates transcription from synthetic promoters termed PaPadR harboring a hexameric OPadR operator upstream of a minimal human cytomegalovirus immediate early promoter (PhCMVmin; Fig. 1A). However, in the presence of FA, aPadR dissociates from its promoter PaPadR, and transgene expression is deactivated.

(A) Schematic design of the FAROFF switch. The binding domain PadR was fused to the transactivation domain VP64 to create an FA-dependent transactivator aPadR (VP64-PadR), which is driven by the simian virus 40 promoter. In the absence of FA (FA), aPadR binds to a chimeric target promoter PaPadR [(OPadR)6-PhCMVmin] and activates the expression of reporter gene SEAP. In the presence of FA (+FA), aPadR is released from PaPadR, and SEAP expression is switched off. (B to D) Dose-dependent SEAP production profile of the FAROFF switch. SEAP expression of HEK-293 cells cotransfected with pLS125 and pLS74 was assessed after cultivation with different concentrations of FA (B), SF (C), or SF tablets (D). (E) FA-induced SEAP expression in different mammalian cell lines. (F) Time-dependent SEAP expression kinetics of the FAROFF switch. SEAP levels of stable cell line HEKFAR-OFF-SEAP, exposed to 500 M SF for different time periods (0 to 72 hours), were profiled at specific time points (X axis, 6 to 72 hours). (G) Reversibility of the FAROFF switch. SEAP expression of HEKFAR-OFF-SEAP cells, alternating the SF concentrations (500 M: OFF; 0 M: ON) every 2 days, was profiled every 12 hours. Data are means SD; n = 3 independent experiments.

To optimize FA/SF-inducible OFF switch (FAROFF switch) characteristics with regard to maximal transgene expression in the absence of FA and minimal basal expression in its presence, we constructed and tested four different configurations of the hybrid transactivator aPadR (fig. S1), four different linkers (La) between VP64 and PadR (fig. S2), and six different tandem repeat variants of OPadR in the synthetic promoters (fig. S3). We found that a combination of aPadR (VP64-La3-PadR) and PaPadR6 [(OPadR)6-PhCMVmin] showed the highest fold induction (~44.1-fold) of the reporter gene SEAP (secreted alkaline phosphatase) when cultivated in the presence or absence of FA (fig. S3F). To exclude that the observed change in gene expression was due to pleiotropic effects of the inducer, we transfected human embryonic kidney (HEK) 293 cells with a constitutive SEAP expression plasmid (pSEAP2-control, PSV40-SEAP-pA) and cultivated the cells in the presence of increasing concentrations (0 to 2000 M) of different inducers (FA, SF, and commercial SF tablets) for 48 hours. Subsequent scoring of SEAP production and cell viability demonstrate that the tested inducers do not have negative effects neither on overall gene expression capacity of the transfected cells (fig. S4, A to C) nor on cell viability within the tested concentration range (fig. S4, D to F).

Next, we investigated gene expression kinetics as function of the inducer concentration. Cotransfection of the constitutive aPadR expression vector pLS125 (PhCMV-VP64-La3-PadR-pA) and the response plasmid pLS74 (PaPadR6-SEAP-pA) encoding a PaPadR6-driven SEAP expression unit resulted in high SEAP production (332.98 12.96 U/liter) (Fig. 1B). Cultivation in the presence of increasing concentrations (0 to 1000 M) of FA, SF, or SF tablets, however, resulted in a dose-dependent decrease in SEAP production up to complete deactivation (Fig. 1, B to D). To assess cross-cell compatibility of FAROFF, we tested the switch in various commonly used mammalian cell lines. The data showed that FAROFF-controlled transgene expression was functional in all cell lines tested (Fig. 1E). Differences in expression level between the different cell lines can be attributed to cell-specific factors such as differences in transfection efficiency as also observed in previous studies (18, 19).

To study long-term performance of the FAROFF switch, we generated and randomly selected eight FAROFF-transgenic stable cell lines (HEKFAR-OFF-SEAP) by transfection of pLS168 (ITR-PaPadR6-SEAP-pA::PhCMV-VP64-La3-PadR-P2A-puromycin-pA-ITR) into HEK-293 cells and clonal selection using the Sleeping Beauty transposon system (36). All the selected cell lines showed SF-induced SEAP repression (fig. S5A), but with different regulation performance, probably as a result of differences in transgene copy number and integration sites that remains beyond control using traditional transient transfection method (19). The cell line with the highest regulation performance was used in follow-up experiments. This cell line HEKFAR-OFF-SEAP showed excellently adjustable gene expression time courses and full reversibility of transgene expression (Fig. 1, F and G). Furthermore, FA-dependent expression performance was observed to be stable for 1 week in cell culture plate (fig. S5B). This comprehensive characterization demonstrating excellent FA-adjustable regulation of gene expression, cross-cell line compatibility, and long-term stability and reversibility strongly suggests that this control switch has a broad application potential.

In certain configurations such as in whole animals, FA/SF-inducible gene expression might be preferable compared to the OFF-type switch characteristics as demonstrated above. To obtain an FA-controlled ON switch (FARON switch) in mammalian cells, we constructed a synthetic mammalian transcription repressor iPadR (KRAB-PadR) by fusing the human Krppel-associated box (KRAB) domain to the N terminus of PadR. The transrepressor iPadR binds and silences constitutive gene expression from synthetic promoters (PiPadR) comprising the simian virus 40 promoter (PSV40) fused to different configurations of OPadR-binding sites. However, in the presence of FA, iPadR dissociates from PiPadR, resulting in the derepression of SEAP production (Fig. 2A). To engineer an optimal FARON switch with maximal transgene expression in the presence of FA and minimal basal expression in its absence, we optimized the FARON switch by testing multiple configurations of OPadR motifs in the promoter (fig. S6) and different linkers (Li) between KRAB and PadR (fig. S7). We found that a combination of the transrepressor iPadR (KRAB-Li6-PadR) and the promoter PiPadR5 [OPadR-PSV40-OPadR(RC)] showed the highest fold induction (~43.0-fold) in the presence of FA (Fig. 2B).

(A) Schematic design of the FARON switch. PadR was fused to a trans-silencing domain KRAB to obtain the FA-dependent transrepressor iPadR (KRAB-PadR) driven by the constitutive simian virus 40 promoter. In the absence of FA (FA), iPadR binds to a chimeric target promoter PiPadR [OPadR-PSV40-OPadR(RC)] and represses PSV40-driven SEAP expression. In the presence of FA (+FA), iPadR is dissociated from the target promoter PiPadR and derepresses SEAP expression. (B to D) Dose-dependent SEAP expression profile of the FARON switch. SEAP expression of HEK-293 cells cotransfected with pLS163 and pLS64 was assessed after cultivation with different concentrations of FA (B), SF (C) or SF tablets (D). (E) Dose-dependent SEAP expression kinetics of the stable cell line HEKFAR-ON-SEAP. (F) Time-dependent SEAP expression kinetics of the FARON switch. SEAP levels of HEKFAR-ON-SEAP cells, incubated with 500 M SF for different time periods (0 to 72 hours), were profiled at different time points (X axis, 6 to 72 hours). (G) Reversibility of the FARON switch. SEAP expression of HEKFAR-ON-SEAP cells, alternating the SF concentrations (500 M: ON; 0 M: OFF) every 2 days, was profiled every 12 hours. Data are means SD; n = 3 independent experiments.

Next, we characterized the FARON switch in detail. We cotransfected the plasmids pLS163 (PSV40-KRAB-Li6-PadR-pA) and pLS64 (PiPadR5-SEAP-pA) into HEK-293 cells and incubated the cells in the presence of different concentrations (0 to 1000 M) of FA, SF, or SF tablets. Scoring SEAP production after 48 hours revealed tight repression in the absence of inducers and a dose-dependent increase in transgene expression correlating with higher inducer concentrations (Fig. 2, B to D). Similarly to the FAROFF (Fig. 1E), the FARON switch switch showed excellent switching properties in various mammalian cell lines, indicating its broad application potential (fig. S8A).

To evaluate long-term expression, adjustability, and reversibility of the FARON switch, we created a HEK-293based stable cell line for FARON-mediated SEAP expression (HEKFAR-ON-SEAP) using the Sleeping Beauty transposon system (36) and subsequent clonal selection. All the selected clones showed significant SF-induced transgene expression with different maximum SEAP production levels (fig. S8B). One of the clones with the best performance (~240-fold induction) out of the eight evaluated ones was used for the following studies (fig. S8B). Characterizing the stable cell line revealed that HEKFAR-ON-SEAP cell line revealed that SEAP expression could be dose-dependently activated for 6 days (Fig. 2E). Furthermore, fine-tunable, inducer-dependent dynamic expression characteristics (Fig. 2F) and excellent reversibility (Fig. 2G) qualify the FARON system as a precise, robust, and adjustable FA/SF-triggered control.

The CRISPR technology is well known for gene editing and genome/epigenome engineering (37). The ability to control the activity of CRISPR with side effectfree inducers will enable tight regulation of arbitrary endogenous genes for precision gene therapy and or for studying the dynamics of transcriptional regulation. Capitalizing on the strong activation and finely adjustable regulation potential of the FARON switch, we engineered three SF-controlled CRISPR-Cas9 systems for activation (PadRa), inhibition (PadRi), and deletion (PadRdel) of endogenous genes in human cells.

We first explored epigenetic remodelingbased activation of endogenous genes (PadRa) (Fig. 3A). In this design, we used the PiPadR6 promoter to promote SF-inducible expression of an engineered activation mediator complex MS2-P65-HSF1 (38). This complex harbors a transcription activation domain (P65) and further an MS2-binding site, which allows its recruitment to dCas9 via single guide RNAs (sgRNAs) fused to MS2-specific RNA aptamers. Thus, the SF-inducible expression of MS2-P65-HSF1 triggers transcription of dCas9-targeted exogenous and endogenous genes. Functionality of this system was successfully validated by the SF-mediated dose-dependent expression of exogenous SEAP gene (Fig. 3B) and endogenous ASCL1, IL1RN, and RHOXF genes (Fig. 3C).

(A) Schematic design of FA-controlled PadR-mediated activation (PadRa). In the presence of SF, the transactivator MS2-P65-HSF1 is produced and recruited by constitutively expressed dCas9 and sgRNAMS2 (sgRNA with MS2 loop) to form a transcriptional activation complex (sgRNAMS2-dCas9-MS2-P65-HSF1). (B) Dose-dependent SF-inducible SEAP expression for PadRa. (C) Dose-dependent PadRa-mediated activation of different endogenous genes, including ASCL1, IL1RN, and RHOXF. (D) Schematic design of SF-controlled PadR-mediated inhibition (PadRi). In the presence of SF, the expression of sgRNA is induced to allow assembly of a repressive complex (sgRNA-dCas9-KRAB) targeting sgRNA-specific DNA sites to inhibit gene expression. (E) Dose-dependent SF-repressible SEAP expression by PadRi. (F) PadRi-mediated repression of different endogenous genes, including CD71 and CXCR4. (G) Schematic design of FA-controlled PadR-mediated gene deletion (PadRdel). Cas9 expression is induced by SF and enables Cas9-mediated target gene deletion. (H) FA-induced gene editing of exogenous gene d2EYFP. The data [(B), (C), (E), (F), and (H)] represent the means SD; n = 3 independent experiments. (I) PadRdel-mediated genome CCR5 and CXCR4 editing. Red arrowheads indicate the expected cleavage bands. N.D.: not detectable; bp: base pair. See table S1 for detailed description of genetic components and table S6 for detailed transfection mixtures.

We next applied the FARON switch for the SF-dependent inhibition of target genes (PadRi) by controlling sgRNA expression to induce the transcriptional repressor dCas9-KRAB to the desired genome region. To this aim, OPadR-binding sites were sandwiched upstream and downstream of the U6 (constitutive) promoter [termed OPadR-PU6-OPadR(RC)] that drives sgRNA expression. When cotransfecting this system with an expression plasmid for iPadR, sgRNA expression is silenced, whereas the addition of SF derepresses U6-driven sgRNA expression by disrupting iPadR binding (Fig. 3D). The produced sgRNA is assembled into a repressive complex (sgRNA-dCas9-KRAB) to target specific DNA sites and to inhibit gene expression. To test whether the PadRi system can achieve sgRNA-mediated target gene inhibition, we cotransfected HEK-293 cells with the PadRi components together with the sgRNAs targeting defined genes. With this system, we used SF to control the expression of exogenous SEAP (Fig. 3E) and d2EYFP (fig. S9A) or of the endogenous CD71 and CXCR4 genes (Fig. 3F).

Last, we repurposed the SF-controlled CRISPR-Cas9 system for controllable gene editing (PadRdel). Cas9-mediated cleavage of an endogenous genomic locus induces indel mutations via nonhomologous end joining (NHEJ), thus disrupting functional gene expression. We used the FARON system with the PiPadR6 promoter to express Cas9 in an SF-inducible manner (Fig. 3G). We first validated functionality of the system by disrupting expression of the exogenous d2EYFP gene in HEK-293 as demonstrated by flow cytometry analysis and by fluorescence microscopy (Fig. 3H and fig. S9B). We next demonstrated the applicability of the PadRdel system to edit the gene of the endogenous CCR5 chemokine receptor that acts as co-receptor for HIV uptake and the c-x-c motif chemokine receptor 4. As detected by the mismatch-sensitive T7 endonuclease I (T7E1) assay, the PadRdel system successfully edited the CCR5 and CXCR4 genes (Fig. 3I).

Mammalian cells engineered with programmable control networks performing higher-order arithmetic calculations may enable the assembly biocomputers to allow the design of complex human-machine interfaces and provide patient-tailored therapeutic interventions in future gene- and cell-based precision therapies (39, 40). By rational combination of mammalian synthetic gene switches, Boolean logic gates could be constructed to program complex intracellular computation that allows cells to process with a computer-like algorithm (15, 18). One of the long-term goals of these efforts is to develop programmable logic computations controlled by side effect-free inputs. To seek a generalizable biocomputing platform controlled by safe, food-derived, and clinically approved molecules, we combined our FARON system with a previously reported system responsive to the food additive BA (15). To demonstrate the general applicability of this approach, we have constructed five examples of logic gates to achieve a variety of Boolean operations controlled by SF and BA. Of special interest are the negation of the OR gate (NOR) and the complement of an AND gate (NAND) operations that are functionally complete, i.e., these operations can be used to express all possible truth tables by combining members of the set into a Boolean expression (41).

First, two BA-controlled genetic switches (BAON and BAOFF) were constructed and validated as reported (15). An AND logic gate is exclusively induced in the presence of both inputs. To engineer an AND gate, we constructed a synthetic promoter PBF1 [PSV40-OPadR-(OCbaR)2] comprising an SF-repressible promoter (PSV40-OPadR) fused to a BA-inducible CbaR-binding site (OCbaR)2. Only in the presence of both SF and BA, the transrepressors KRAB-PadR and KRAB-CbaR are released and derepress expression of the reporter d2EYFP (Fig. 4A). As a complementary phase to the AND gate, a NAND gate was designed. To this aim, we constructed a chimeric promoter PBF2 [(OCbaR)2-OPadR-PhCMVmin] comprising an SF-inducible promoter (OPadR-PhCMVmin) fused to a CbaR-binding site (OCbaR)2 in front of the OPadR site. Only in the presence of both SF and BA, the transactivators VP16-CbaR and VP64-PadR dissociate from their cognate operators, resulting in effective deactivation of d2EYFP expression (Fig. 4B).

Processing performance of the (A) AND, (B) NAND, (C) NOR, (D) SF NIMPLY BA, and (E) BA NIMPLY SF logic gates in HEK-293 cells. The schematic design of the five logic gates is displayed on the left panels. The processing output performance of five logic gates is shown on the right panels. HEK-293 cells (8 104) were cotransfected with (A) pLS25, pMX34, and pDL55; (B) pLS56, pMX43, and pDL58; (C) pLS125, pDL41, and pDL68; (D) pMX43, pLS25, and pDL73; and (E) pLS125, pMX34, and pDL77 and cultivated with different combinations of the two input signals SF (1 mM) and BA (0.75 mM) in accordance with the truth tables. d2EYFP expression in the cells was quantified by fluorescence microscopy and flow cytometric analysis 48 hours after the addition of input signals. Data are means SD; n = 3 independent experiments. See table S1 for detailed description of genetic components and table S7 for detailed transfection mixtures for each logic gate.

A NOR gate is turned on only when no input signal is present. A NOR gate was constructed by wiring the FAROFF and BAOFF switches in a cascade configuration. In the absence of both inducers, SF and BA, the constitutively expressed transactivator VP64-PadR binds to the SF-responsive minimal promoter PaPadR6 [(OPadR)6-PhCMVmin] to initiate expression of the BA-responsive transactivator CbaR-VP16, which, in turn, binds to the BA-responsive minimal promoter PBF3 [(OCbaR)2-PhCMVmin] to induce expression of the output gene d2EYFP (Fig. 4C). An A NIMPLY B gate is exclusively induced in the presence of A while B is absent (42). For example, in an SF NIMPLY BA gate, the output is expected to be only induced if SF is present while BA is absent. To engineer the SF NIMPLY BA logic gate, we assembled a chimeric promoter PBF4 [(OCbaR)2-PhCMVmin-OPadR] comprising a BA-deactivatable promoter [(OCbaR)2-PhCMVmin] flanked in 3 by a PadR-binding site OPadR. Only in the presence of SF and the absence of BA, the transrepressor KRAB-PadR dissociates from PBF4, while the transactivator CbaR-VP16 binds to PBF4 to activate d2EYFP expression (Fig. 4D). To obtain a reversed BA NIMPLY SF logic gate, we engineered a chimeric promoter PBF5 [(OPadR)6-PhCMVmin-(OCbaR)2] comprising an SF-deactivatable promoter [(OPadR)6-PhCMVmin] flanked in 3 by a CbaR-binding site (OCbaR)2. Only in the presence of BA and the absence of SF, the transrepressor KRAB-CbaR dissociates from PBF5, while the transactivator VP64-PadR binds to PBF5 to activate d2EYFP expression (Fig. 4E). These exciting proof-of-concept results for biocomputing show that complex genetic programs could precisely be controlled by food additives and clinically licensed drugs, indicating the application potential for complex engineered cell-based therapeutic dosing regimens.

For future applications in precision gene- and cell-based therapies, it is essential that state-of-the-art gene regulation systems are functional within whole organisms (43). To validate SF-controlled transgene gene expression in animals, we microencapsulated HEKFAR-ON-SEAP cells into coherent, immunoprotective alginate-poly-(l-lysine)-alginate beads. After confirming SF-inducible SEAP production in microencapsulated cells in vitro (fig. S10), we next implanted the cell-containing capsules intraperitoneally into mice (Fig. 5A). The mice were given a dose of SF within the range of 1 to 1000 mg/kg each day. Notably, the acute LD50 (median lethal dose) of 3200 mg/kg was calculated in mice (35). SEAP production quantified in the blood showed SF-dependent dose-response kinetics for up to 15 days (Fig. 5B). When the mice were implanted with HEKFAR-ON-SEAP and orally administrated with clinically approved solubilized SF tablets, we observed significantly induced SEAP levels in the bloodstream for 15 days as well (Fig. 5C). However, we observed SEAP levels in vivo decreased at day 15, which may be attributed to the cell encapsulation technologies containing xenogeneic molecules that may escape from capsules and trigger immune responses destroying the encapsulated cells or their surrounding tissues to hinder the long-term performance of HEKFAR-ON-SEAP cells (22). Despite the limitations of transplant technology, these results demonstrate that SF tablets can be used as a trigger for the tight and precise control of implanted cells in future clinical investigations.

(A) Schematic showing the mouse experimental design and procedure for assessing SF-controlled transgene expression in vivo. HEK-293 cells engineered for FARON-inducible SEAP production were microencapsulated into alginate-poly-(l-lysine)-alginate beads allowing free diffusion of oxygen, nutrients, and secreted proteins across the membrane while simultaneously shielding encapsulated cells from the immune system. SEAP production in microencapsulated HEKFAR-ON-SEAP cells implanted in the peritoneum of mice could be controlled either via injection of SF or via oral administration of SF tablets. (B and C) Dose-dependent SF-inducible SEAP expression in mice. Eight-week-old male C57BL/6J mice were intraperitoneally implanted with 2 106 microencapsulated HEKFAR-ON-SEAP cells (200 cells per capsule) and received intraperitoneal administration of SF (B) or oral administration of clinically licensed SF tablets (C) three times per day (0 to 1000 mg/kg per day). SEAP levels in the bloodstream of mice were quantified on days 2, 4, 8, and 15 after implantation. Data are expressed as means SEM; n = 5 to 6 mice. n.s., not significant. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 versus control.

In this work, we have developed an FA/SF-controlled transcription switch (FAR switch), which can be applied to control transgene expression, genome/epigenome engineering, and complex biological logic operations in mammalian cells. One of the great advantages of the FAR switch is that it is responsive to an off-the-shelf clinical drug that was shown to be safe and does not trigger inadvertent side reactions (31). SF has been used to treat patients with cardiovascular and cerebrovascular diseases for decades in China (31), indicating that SF is very safe for long-term therapeutic usage. Such beneficial properties of the inducer might facilitate the clinical approval for the corresponding gene switch. Another potential benefit of SF being clinically licensed is the perspective of developing a combined therapy. In such a perspective, SF not only would coordinate both the expression of desired therapeutic genes but would also elicit its original therapeutic effect. For example, the system could be deployed to control both expression of antithrombotic peptides expression, such as hirudin (44) or antithrombotic bioactive peptides (45), and the intrinsic antithrombotic activity of SF. Such scenarios would represent an attractive therapeutic option as the original activity of a drug could be invigorated and improved by a complementing therapeutic gene network.

Combining the PadR repressor from B. subtilis 168 (35) and mammalian epigenetic regulators, we have constructed the novel mammalian transgene expression systems FARON and FAROFF, which respond to the clinically approved drug SF. The optimized FARON/FAROFF switch demonstrated very low background, high induction folds, robust induction kinetics, precise adjustability, and full reversibility. Because of the high modularity of the switch, we were able to construct SF-controlled Cas9/dCas9 systems for controllable genome editing and epigenome remodeling. Moreover, the FAR switch showed interference-free regulation characteristics when used with the BA-inducible switch, thus enabling to perform logic computations in mammalian cells, indicating that this switch is likely an ideal building block for the design of complex and high-order synthetic gene networks. The suitability of the FARON to regulate gene expression in mice opens the perspective of transferring such complex regulation networks into a therapeutic setting functional in living mammals.

Despite the strong regulation performance in vitro and in vivo, there is room for improvement with regard to a translational perspective. For example, the sensitivity of the FAR switch to its inducer could further be improved to obtain a higher dynamic range in vivo in response to SF tablets. We anticipate that the sensitivity of the FAR switch could be optimized by engineering the transcription factor and the operator sequence in the promoter. For example, by using directed evolution approaches (46) or rational protein design based on bioinformatics (47), it may be possible to obtain a more sensitive mutated version of the transcriptional repressor PadR.

Collectively, our work demonstrates how genetic switches responsive to the clinically approved drug SF can expand the synthetic biology tool box and provide a new device to cybergenetics. This work shows a safe and robust strategy for dynamic control of transgene expression and epigenetic engineering, which provides an alternative strategy to achieve dynamic interventions in future gene- and cell-based precision medicine.

Comprehensive design and construction details for all expression vectors are listed in table S1. The expression vectors were constructed by T4 DNA ligation (Takara Bio, catalog no. 2011B) or Gibson assembly according to the manufacturers instructions [Seamless Assembly Cloning Kit, Obio Technology Inc., catalog no. BACR(C) 20144001]. All constructions have been confirmed by sequencing (Genewiz Inc., China).

Human cervical adenocarcinoma cells [HeLa, American Type Culture Collection (ATCC): CCL-2], HEK cells (HEK-293, ATCC: CRL-11268), HEK-293derived Hana3A cells that were engineered for the stable expression of G and chaperones RTP1/RTP2/REEP1, and telomerase-immortalized human mesenchymal stem cells (hMSC-TERT, ATCC: SCRC4000) were cultivated in Dulbeccos modified Eagles medium (DMEM; Gibco, lot no. 31600-083) supplemented with 10% (v/v) fetal bovine serum (FBS; Biological Industries, lot no. 04-001-1C) and 1% (v/v) penicillin/streptomycin solution. Chinese hamster ovary cells (CHO-K1, ATCC: CCL-61) were cultivated in its specific culture medium (Procell, lot no. CM-0062) supplemented with 5% (v/v) FBS and 1% (v/v) penicillin/streptomycin solution. All cell types were cultured at 37C in a humidified atmosphere containing 5% CO2.

All cell lines were transfected with a standard polyethyleneimine (PEI)based protocol. Briefly, 5.0 104 HEK-293 cells were seeded in a 24-well cell culture plate. After 20 hours of cultivation, 200 ng of plasmid DNA diluted in 50 l of FBS-free DMEM was mixed with 0.6 l of PEI (PolyScience, catalog no. 24765; 1 g/l). The transfection mixture was incubated at room temperature for 15 min and subsequently dropwise added to the cells. After 6 hours, the culture medium was replaced by fresh cell culture medium optionally supplemented with different concentrations of FA (Sigma-Aldrich, catalog no. 1270311), SF (Qufu Hongly Chemical Industry Co. Ltd., catalog no. 24276-84-4), or SF tablets (Chengdu Hengda Pharmaceutical Co. Ltd., guoyaozhunzi: H51023583). Cell concentrations and viability of all cell lines were profiled with a Countess II Automated cell counter (Life Technologies, USA).

To test the impact of various types of inducers (FA, SF, and SF tablets) on human cells, 1 104 HEK-293 cells per well were seeded in a 96-well plate, transfected with the reporter plasmid pSEAP2-control (PSV40-SEAP-pA; 50 ng), and incubated with different concentrations of chemicals (0 to 2000 M) for 48 hours. The cell culture supernatant was collected for the quantification of SEAP expression (48).

To test the effects of inducers on cell viability, 1 104 HEK-293 cells per well were seeded in a 96-well plate and incubated with different concentrations of inducers (0 to 2000 M) for 48 hours. Then, the cell culture medium was replaced by 100 l of fresh medium containing 10 l of an [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] (MTT) solution (5 mg/ml; Sangon Biotech, catalog no. A600799) and incubated for 4 hours at 37C. After incubation, the medium was replaced with 100 l of dimethyl sulfoxide (Sangon Biotech, catalog no. A600163) and incubated for 10 min at 37C. The light absorbance was measured at 540 nm using a Synergy H1 hybrid multimode microplate reader (BioTek Instruments Inc.) using Gen5 software (version 2.04).

Human placental SEAP in cell culture medium was quantified using a p-nitrophenylphosphatebased light absorbance time course assay, as described previously (18). The SEAP production in mice was quantified with a chemiluminescence-based assay kit (Roche Diagnostics, catalog no. 11779842001) according to the manufacturers protocol.

The Sleeping Beauty (SB) transposon system was used for selecting stable cell lines as described before (36). The HEKFAR-ON-SEAP cell line, transgenic for SF-inducible SEAP expression, was constructed by cotransfecting HEK-293 cells with pLS180 [ITR-PiPadR5-SEAP-pA::PhCMV-KRAB-Li6-PadR-P2A-puromycin-pA-ITR; 200 ng] and the Sleeping Beauty transposase expression vector PhCMV-T7-SB100 (PhCMV-SB100X-pA; 20 ng) and selected with puromycin (1 g/ml) for 2 weeks. The living cells were picked for further cultivation and induced with 1 mM SF for 48 hours. The monoclonal HEKFAR-ON-SEAP cell line showing the highest fold induction in gene expression in response to SF was used for subsequent studies.

The genomic DNA of treated HEK-293 cells was extracted using a TIANamp Genomic DNA Extraction Kit (TIANGEN Biotech Inc., catalog no. DP304) according to the manufacturers protocol. Then, sgRNA-targeted CCR5 genes were polymerase chain reaction (PCR)amplified from genomic DNA using the primers listed in table S2. The PCR amplicons were purified from agarose gel with a Universal DNA Purification Kit (TIANGEN Biotech Inc., catalog no. DP214). Twenty microliters of the reaction mixture containing 500 ng of purified PCR products and 2 l of 10 M buffer (TIANGEN Biotech Inc., catalog no. 1060S) were reannealed (95C for 5 min, and then slowly cooled to room temperature) to form heteroduplex DNA. Then, 0.5 l of T7 endonuclease I (New England Biolabs, catalog no. M0302) was added and incubated at 37C for 2 hours. The digested products were analyzed by 2% agarose gel electrophoresis. The percentage of deletion by Cas9 was calculated with the following formula: 100% (b + c)/(a + b + c), where a represented the intensity of undigested PCR amplicons and b and c represented the intensities of the T7E1-digested products.

Total RNA of treated HEK-293 cells was isolated using the RNAiso Plus Kit (Takara Bio, catalog no. 9108). One microgram of purified RNA was reverse-transcribed into complementary DNA (cDNA) using a PrimeScript Reverse Transcription Kit with the gDNA Eraser (Takara Bio, catalog no. RR047). Quantitative PCR (qPCR) was performed by SYBR Premix Ex Taq (Takara Bio, catalog no. RR420) with primers listed in table S3 and profiled by QuantStudio (Thermo Fisher Scientific Inc.) with the following amplification parameters: 95C for 10 min, 40 cycles of 95C for 30 s, 60C for 30 s, and 72C for 30 s, and a final extension at 72C for 10 min. All samples were normalized against the endogenous housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and the results were evaluated as the relative mRNA level using the standard Ct method (49).

The expression of d2EYFP was visualized using an Olympus microscope (Olympus IX71, TH4-200) equipped with an Olympus digital camera, a pE-100-LED (CoolLED) as the transmission light source, a Spectra X (Lumencor) as the fluorescent light source, a 10 objective, a 488-nm/509-nm (Blue/Green/Red) excitation/emission filter set, and the Image-Pro Express C software (version ipp6.0). Identical settings including exposure times for d2EYFP were used for all fluorescence micrographs.

Flow cytometric analysis was performed as described previously (18). In brief, cells were analyzed with a Becton Dickinson FACSCalibur Flow Cytometer (BD Biosciences) equipped for d2EYFP [488-nm laser, 505-nm long pass filter, 530/30 emission filter (passband centered on 530 nm; passband width: 30 nm)] detection and set to exclude dead cells and cell doublets. About 10,000 cells were recorded in each dataset and analyzed with the BD CellQuest Pro software (version no. 6.0). To quantify the expression profiles of the FA-controlled gene deletion and SF/BA-controlled programmable biocomputers in human cells, transfected HEK-293 populations were gated for cells with high d2EYFP fluorescence beyond a threshold of 103 arbitrary fluorescence units. The percentage of gated cells was multiplied by their median fluorescence, resulting in a weighted d2EYFP expression profile that correlated fluorescence intensity with cell number.

The HEKFAR-ON-SEAP stable cell line engineered for SF-controlled SEAP expression was encapsulated into coherent alginate-poly-(l-lysine)-alginate beads (400-m diameter; 200 cells per capsule) using a B-395 Pro Encapsulator (BUCHI Labortechnik AG) according to the manufacturers instructions. Microcapsules were generated with the following parameters: 200-m nozzle with a vibration frequency of 1300 Hz, 20-ml syringe operated at a flow rate of 450 U, and 1.10 kV for bead dispersion. Eight-week-old male wild-type C57BL/6 J mice [East China Normal University (ECNU) Laboratory Animal Centre] were intraperitoneally injected with 2 106 microencapsulated HEKFAR-ON-SEAP cells. The mice were then intraperitoneally injected daily with SF or orally administrated with SF tablets at doses ranging from 0 to 1000 mg/kg. Blood samples were collected, and SEAP levels were quantified by a chemiluminescence-based assay kit. The serum was isolated at the indicated points in time after implantation clotting of the blood (37C for 1 hour and then 4C for 1.5 hours) and subsequent centrifugation (2700g, 10 min).

All experiments involving mice were performed according to the directives approved by the ECNU Animal Care and Use Committee (protocol ID: m20140301). All the procedures for samples or data collection used were carried out in compliance with the Ministry of Science and Technology of the Peoples Republic of China on Animal Care Guidelines. All mice were euthanized after the experiments.

All in vitro data represent means SD of three independent experiments (n = 3). For mouse experiments, each treatment group was composed of five to six mice (n = 5 to 6). The blood sample analysis was blinded. Comparisons between groups were analyzed using Students t tests, and the values are expressed as means SD. Differences were considered statistically significant at P < 0.05. Prism 6 software (version 6.01, GraphPad Software Inc.) was used for statistical analysis.

Acknowledgments: We thank the ECNU Multifunctional Platform for Innovation (011) for supporting the mouse experiments and the Instruments Sharing Platform of School of Life Sciences at ECNU for supporting research. We also thank C. Huang for help with the artwork. Funding: This work was financially supported by grants from the National Key R&D Program of China, Synthetic Biology Research (no. 2019YFA0904500), the National Natural Science Foundation of China (NSFC: nos. 31971346 and 31861143016), the Science and Technology Commission of Shanghai Municipality (no. 18JC1411000), the Thousand Youth Talents Plan of China, and the Fundamental Research Funds for the Central Universities to H.Y. This work was also partially supported by NSFC: no. 31901023 to N.G. W.W. was supported by the Deutsche Forschungsgemeinschaft (DFG; German Research Foundation) under Germanys Excellence StrategyEXC-2189Project ID: 390939984. Author contributions: H.Y. conceived this study. H.Y., Y.W., and S.L. designed the project. Y.W., S.L., Y.L., and K.D. performed the experimental work. Y.W., S.L., and K.D. performed the mouse experiments. H.Y., N.G., W.W., and Y.W. analyzed the results and wrote the manuscript. All authors read and approved the manuscript. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. All genetic components related to this paper are available with a material transfer agreement and may be requested from H.Y. (hfye{at}bio.ecnu.edu.cn).

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A versatile genetic control system in mammalian cells and mice responsive to clinically licensed sodium ferulate - Science Advances

Have your cake and eat it too, so long as they’re GMO: 5 Reasons GMO should be on your shopping list – SynBioBeta

Impossible Burger is a household name best recognized for its successful introduction of a plant-based burger that bleeds and has no animal hormones or antibiotics. But, there is another notable first for Impossible Burgerit is the first product in stores to adopt the new USDA-approved bioengineered GMO (genetically modified organism) product label that will soon be federally-mandated. This puts a spotlight on a major debate: the safety of GMO products.

While there have been no proven documented cases of GMOs causing harm in humans or animals, numerous studies show that consumers generally dont understand or lack familiarity with GMOs and are decidedly wary to try products that are genetically engineered.

For example, the pro-GMO organization GMO Answers found that 70% of adults dont really know what GMOs are, and less than a third are comfortable having GMOs in their food. While nearly half of U.S. consumers say they would change their consumption habits to reduce their impact on the environment, its worth noting that organic and natural certifications are anti-GMO, creating an increasingly significant dilemma. Plus a new Regenerative Organic Certification adds soil health, animal welfare and farmer economics to organics, but remains unappreciative that GMOs are a major lever to achieve these new goals. While modern genetic engineering is a key enabler of sustainability, food security and health, many consumers automatically look at GMO labels unfavorably.

There continues to be a serious divide between the scientific community and consumer audiences, certifying that authorities are failing to mediate properly and consumers are receiving an inadequate education on the pros and cons of GMOs. As a hero technology for sustainability, nourishing the population, and supporting farmer economics, GMOs fall victim to a misguided negative perception and are actually quite aligned with prevailing consumer demands.

Here are five considerations to put myths to rest and make the case for GMOs:

A GMO has had its DNA altered or modified in some way through genetic engineering. Approximately 60% of all processed foods on supermarket shelves contain GMO ingredients and, according to the USDA, 94% of soy and 92% of corn grown in the United States is GMO. Additionally, more than 90% of the corn and soy harvested for feed utilized in the production of livestock is GMO.

Humans have been altering the genetic makeup of plants for millennia, keeping seeds from the best crops and planting them in following years, selective breeding and crossbreeding, and conducting induced mutation to enable new varieties of crops that taste sweeter, grow bigger, and last longer. It is the technique of genetic-engineering that is newmodern genetic engineering utilizes biotechnology to intentionally direct a targeted change in a plant, animal, or microbial gene sequence to achieve a specific result.

Take for instance watermelon. The watermelons we eat today contrast starkly with those depicted in a 17th-century painting by Italian artist Giovanni Stanchi. Over time, genetic modification through selective breeding has enabled watermelons of more consistent shape, fewer seeds, increased water and sugar, and bright, red flesh. Which version would you rather eat? Chances are, youd choose the modern variety, which is also the more profitable type for the farmer and higher performing in terms of yield and nutrition.

GMO products undergo more rigorous testing than other foods we consume and are screened for toxins, allergens, nutrients, and proteins to make sure they are safe for human consumption. Additionally, Hundreds of digestion and safety studies examining the effects of feeding genetically engineered crops to various food-producing animal species revealed no disturbance to nutritional value, quality, or health. Regarding environmental safety, GMOs enable decreased reliance on chemical sprays that are harmful to soil and water run-off, and are controlled to mitigate gene flow.

Our food system has reached a pivotal moment. The United Nations estimates our global population will reach 9.7 billion by 2050, all of whom will depend on access to safe, nourishing, and affordable food, which the system today cannot support due to limited land, water scarcity, disease resistance, and climate change. If we are to dramatically improve production, we need to be able to improve yields across current acreage, expand farming in new regions and soil types, grow crops that can tolerate destructive weeds, pests, and molds, and perform in a changing climate. Next-gen biotechnology allows us to do this with greater success, speeds, safety and leads to novel breakthroughs in sustainable methods of production.

820 million people globally are malnourished and biotechnology is the best toolset in our arsenal for addressing the urgent problems of food shortage and hunger, globally. As an example, a 1995 report by the World Health Organization, estimated that more than 254 million children of preschool ages across 60 countries suffered from vitamin A deficiency, which can lead to permanent blindness and death. In 1999 a team of scientists leveraged GMO-based biofortification to introduce two daffodil genes and one bacterial gene into rice plants that enable the staple crop to produce in its grains beta-carotene, a building block of vitamin A. The result was a genetically-engineered crop carrying a promise to prevent millions of deaths and alleviate the suffering of children and adults afflicted with vitamin A deficiency and micronutrient malnutrition in developing countries.

Consumers are plagued by prolific misinformation throughout the food industry. GMOs are just one variable in a complicated web that includes organic, all-natural, free-range, hormone-free, antibiotic-free, and a cornucopia of others that consumers navigate with varying degrees of accuracy to settle on a food ideology.

As experts and food system stakeholders, the onus is on us to better facilitate the communication between consumers and producers, as well as credibly separate myth from fact while in pursuit of feeding everyone sustainably. Consumers are increasingly shopping with purpose and we need to help them be unconflicted and successful.

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Have your cake and eat it too, so long as they're GMO: 5 Reasons GMO should be on your shopping list - SynBioBeta

Biotechnology Could Change the Cattle Industry. Will it Succeed? – Undark Magazine

When Ralph Fisher, a Texas cattle rancher, set eyes on one of the worlds first cloned calves in August 1999, he didnt care what the scientists said: He knew it was his old Brahman bull, Chance, born again. About a year earlier, veterinarians at Texas A&M extracted DNA from one of Chances moles and used the sample to create a genetic double. Chance didnt live to meet his second self, but when the calf was born, Fisher christened him Second Chance, convinced he was the same animal.

Scientists cautioned Fisher that clones are more like twins than carbon copies: The two may act or even look different from one another. But as far as Fisher was concerned, Second Chance was Chance. Not only did they look identical from a certain distance, they behaved the same way as well. They ate with the same odd mannerisms; laid in the same spot in the yard. But in 2003, Second Chance attacked Fisher and tried to gore him with his horns. About 18 months later, the bull tossed Fisher into the air like an inconvenience and rammed him into the fence. Despite 80 stitches and a torn scrotum, Fisher resisted the idea that Second Chance was unlike his tame namesake, telling the radio program This American Life that I forgive him, you know?

In the two decades since Second Chance marked a genetic engineering milestone, cattle have secured a place on the front lines of biotechnology research. Today, scientists around the world are using cutting-edge technologies, from subcutaneous biosensors to specialized food supplements, in an effort to improve safety and efficiency within the $385 billion global cattle meat industry. Beyond boosting profits, their efforts are driven by an imminent climate crisis, in which cattle play a significant role, and growing concern for livestock welfare among consumers.

Gene editing stands out as the most revolutionary of these technologies. Although gene-edited cattle have yet to be granted approval for human consumption, researchers say tools like Crispr-Cas9 could let them improve on conventional breeding practices and create cows that are healthier, meatier, and less detrimental to the environment. Cows are also being given genes from the human immune system to create antibodies in the fight against Covid-19. (The genes of non-bovine livestock such as pigs and goats, meanwhile, have been hacked to grow transplantable human organs and produce cancer drugs in their milk.)

But some experts worry biotech cattle may never make it out of the barn. For one thing, theres the optics issue: Gene editing tends to grab headlines for its role in controversial research and biotech blunders. Crispr-Cas9 is often celebrated for its potential to alter the blueprint of life, but that enormous promise can become a liability in the hands of rogue and unscrupulous researchers, tempting regulatory agencies to toughen restrictions on the technologys use. And its unclear how eager the public will be to buy beef from gene-edited animals. So the question isnt just if the technology will work in developing supercharged cattle, but whether consumers and regulators will support it.

Cattle are catalysts for climate change. Livestock account for an estimated 14.5 percent of greenhouse gas emissions from human activities, of which cattle are responsible for about two thirds, according to the United Nations Food and Agriculture Organization (FAO). One simple way to address the issue is to eat less meat. But meat consumption is expected to increase along with global population and average income. A 2012 report by the FAO projected that meat production will increase by 76 percent by 2050, as beef consumption increases by 1.2 percent annually. And the United States is projected to set a record for beef production in 2021, according to the Department of Agriculture.

For Alison Van Eenennaam, an animal geneticist at the University of California, Davis, part of the answer is creating more efficient cattle that rely on fewer resources. According to Van Eenennaam, the number of dairy cows in the United States decreased from around 25 million in the 1940s to around 9 million in 2007, while milk production has increased by nearly 60 percent. Van Eenennaam credits this boost in productivity to conventional selective breeding.

You dont need to be a rocket scientist or even a mathematician to figure out that the environmental footprint or the greenhouse gases associated with a glass of milk today is about one-third of that associated with a glass of milk in the 1940s, she says. Anything you can do to accelerate the rate of conventional breeding is going to reduce the environmental footprint of a glass of milk or a pound of meat.

Modern gene-editing tools may fuel that acceleration. By making precise cuts to DNA, geneticists insert or remove naturally occurring genes associated with specific traits. Some experts insist that gene editing has the potential to spark a new food revolution.

The question isnt just if the technology will work in developing supercharged cattle, but whether consumers and regulators will support it.

Jon Oatley, a reproductive biologist at Washington State University, wants to use Crispr-Cas9 to fine tune the genetic code of rugged, disease-resistant, and heat-tolerant bulls that have been bred to thrive on the open range. By disabling a gene called NANOS2, he says he aims to eliminate the capacity for a bull to make his own sperm, turning the recipient into a surrogate for sperm-producing stem cells from more productive prized stock. These surrogate sires, equipped with sperm from prize bulls, would then be released into range herds that are often genetically isolated and difficult to access, and the premium genes would then be transmitted to their offspring.

Furthermore, surrogate sires would enable ranchers to introduce desired traits without having to wrangle their herd into one place for artificial insemination, says Oatley. He envisions the gene-edited bulls serving herds in tropical regions like Brazil, the worlds largest beef exporter and home to around 200 million of the approximately 1.5 billion head of cattle on Earth.

Brazils herds are dominated by Nelore, a hardy breed that lacks the carcass and meat quality of breeds like Angus but can withstand high heat and humidity. Put an Angus bull on a tropical pasture and hes probably going to last maybe a month before he succumbs to the environment, says Oatley, while a Nelore bull carrying Angus sperm would have no problem with the climate.

The goal, according to Oatley, is to introduce genes from beefier bulls into these less efficient herds, increasing their productivity and decreasing their overall impact on the environment. We have shrinking resources, he says, and need new, innovative strategies for making those limited resources last.

Oatley has demonstrated his technique in mice but faces challenges with livestock. For starters, disabling NANOS2 does not definitively prevent the surrogate bull from producing some of its own sperm. And while Oatley has shown he can transplant sperm-producing cells into surrogate livestock, researchers have not yet published evidence showing that the surrogates produce enough quality sperm to support natural fertilization. How many cells will you need to make this bull actually fertile? asks Ina Dobrinski, a reproductive biologist at the University of Calgary who helped pioneer germ cell transplantation in large animals.

But Oatleys greatest challenge may be one shared with others in the bioengineered cattle industry: overcoming regulatory restrictions and societal suspicion. Surrogate sires would be classified as gene-edited animals by the Food and Drug Administration, meaning theyd face a rigorous approval process before their offspring could be sold for human consumption. But Oatley maintains that if his method is successful, the sperm itself would not be gene-edited, nor would the resulting offspring. The only gene-edited specimens would be the surrogate sires, which act like vessels in which the elite sperm travel.

Even so, says Dobrinski, Thats a very detailed difference and Im not sure how that will work with regulatory and consumer acceptance.

In fact, American attitudes towards gene editing have been generally positive when the modification is in the interest of animal welfare. Many dairy farmers prefer hornless cows horns can inflict damage when wielded by 1,500-pound animals so they often burn them off in a painful process using corrosive chemicals and scalding irons. In a study published last year in the journal PLOS One, researchers found that most Americans are willing to consume food products from cows genetically modified to be hornless.

Still, experts say several high-profile gene-editing failures in livestock and humans in recent years may lead consumers to consider new biotechnologies to be dangerous and unwieldy.

In 2014, a Minnesota startup called Recombinetics, a company with which Van Eenennaams lab has collaborated, created a pair of cross-bred Holstein bulls using the gene-editing tool TALENs, a precursor to Crispr-Cas9, making cuts to the bovine DNA and altering the genes to prevent the bulls from growing horns. Holstein cattle, which almost always carry horned genes, are highly productive dairy cows, so using conventional breeding to introduce hornless genes from less productive breeds can compromise the Holsteins productivity. Gene editing offered a chance to introduce only the genes Recombinetics wanted. Their hope was to use this experiment to prove that milk from the bulls female progeny was nutritionally equivalent to milk from non-edited stock. Such results could inform future efforts to make Holsteins hornless but no less productive.

The experiment seemed to work. In 2015, Buri and Spotigy were born. Over the next few years, the breakthrough received widespread media coverage, and when Buris hornless descendant graced the cover of Wired magazine in April 2019, it did so as the ostensible face of the livestock industrys future.

But early last year, a bioinformatician at the FDA ran a test on Buris genome and discovered an unexpected sliver of genetic code that didnt belong. Traces of bacterial DNA called a plasmid, which Recombinetics used to edit the bulls genome, had stayed behind in the editing process, carrying genes linked to antibiotic resistance in bacteria. After the agency published its findings, the media reaction was swift and fierce: FDA finds a surprise in gene-edited cattle: antibiotic-resistant, non-bovine DNA, read one headline. Part cow, part bacterium? read another.

Recombinetics has since insisted that the leftover plasmid DNA was likely harmless and stressed that this sort of genetic slipup is not uncommon.

Is there any risk with the plasmid? I would say theres none, says Tad Sonstegard, president and CEO of Acceligen, a Recombinetics subsidiary. We eat plasmids all the time, and were filled with microorganisms in our body that have plasmids. In hindsight, Sonstegard says his teams only mistake was not properly screening for the plasmid to begin with.

While the presence of antibiotic-resistant plasmid genes in beef probably does not pose a direct threat to consumers, according to Jennifer Kuzma, a professor of science and technology policy and co-director of the Genetic Engineering and Society Center at North Carolina State University, it does raise the possible risk of introducing antibiotic-resistant genes into the microflora of peoples digestive systems. Although unlikely, organisms in the gut could integrate those genes into their own DNA and, as a result, proliferate antibiotic resistance, making it more difficult to fight off bacterial diseases.

The lesson that I think is learned there is that science is never 100 percent certain, and that when youre doing a risk assessment, having some humility in your technology product is important, because you never know what youre going to discover further down the road, she says. In the case of Recombinetics. I dont think there was any ill intent on the part of the researchers, but sometimes being very optimistic about your technology and enthusiastic about it causes you to have blinders on when it comes to risk assessment.

The FDA eventually clarified its results, insisting that the study was meant only to publicize the presence of the plasmid, not to suggest the bacterial DNA was necessarily dangerous. Nonetheless, the damage was done. As a result of the blunder,a plan was quashed forRecombinetics to raise an experimental herd in Brazil.

Sometimes being very optimistic about your technology and enthusiastic about it causes you to have blinders on when it comes to risk assessment.

Backlash to the FDA study exposed a fundamental disagreement between the agency and livestock biotechnologists. Scientists like Van Eenennaam, who in 2017 received a $500,000 grant from the Department of Agriculture to study Buris progeny, disagree with the FDAs strict regulatory approach to gene-edited animals. Typical GMOs are transgenic, meaning they have genes from multiple different species, but modern gene-editing techniques allow scientists to stay roughly within the confines of conventional breeding, adding and removing traits that naturally occur within the species. That said, gene editing is not yet free from errors and sometimes intended changes result in unintended alterations, notes Heather Lombardi, division director of animal bioengineering and cellular therapies at the FDAs Center for Veterinary Medicine. For that reason, the FDA remains cautious.

Theres a lot out there that I think is still unknown in terms of unintended consequences associated with using genome-editing technology, says Lombardi. Were just trying to get an understanding of what the potential impact is, if any, on safety.

Bhanu Telugu, an animal scientist at the University of Maryland and president and chief science officer at the agriculture technology startup RenOVAte Biosciences, worries that biotech companies will migrate their experiments to countries with looser regulatory environments. Perhaps more pressingly, he says strict regulation requiring long and expensive approval processes may incentivize these companies to work only on traits that are most profitable, rather than those that may have the greatest benefit for livestock and society, such as animal well-being and the environment.

What company would be willing to spend $20 million on potentially alleviating heat stress at this point? he asks.

On a windy winter afternoon, Raluca Mateescu leaned against a fence post at the University of Floridas Beef Teaching Unit while a Brahman heifer sniffed inquisitively at the air and reached out its tongue in search of unseen food. Since 2017, Mateescu, an animal geneticist at the university, has been part of a team studying heat and humidity tolerance in breeds like Brahman and Brangus (a mix between Brahman and Angus cattle). Her aim is to identify the genetic markers that contribute to a breeds climate resilience, markers that might lead to more precise breeding and gene-editing practices.

In the South, Mateescu says, heat and humidity are a major problem. That poses a stress to the animals because theyre selected for intense production to produce milk or grow fast and produce a lot of muscle and fat.

Like Nelore cattle in South America, Brahman are well-suited for tropical and subtropical climates, but their high tolerance for heat and humidity comes at the cost of lower meat quality than other breeds. Mateescu and her team have examined skin biopsies and found that relatively large sweat glands allow Brahman to better regulate their internal body temperature. With funding from the USDAs National Institute of Food and Agriculture, the researchers now plan to identify specific genetic markers that correlate with tolerance to tropical conditions.

If were selecting for animals that produce more without having a way to cool off, were going to run into trouble, she says.

A Brahman cow at the University of Floridas Beef Teaching Unit. Visual: Dyllan Furness

There are other avenues in biotechnology beyond gene editing that may help reduce the cattle industrys footprint. Although still early in their development, lab-cultured meats may someday undermine todays beef producers by offering consumers an affordable alternative to the conventionally grown product, without the animal welfare and environmental concerns that arise from eating beef harvested from a carcass.

Other biotech techniques hope to improve the beef industry without displacing it. In Switzerland, scientists at a startup called Mootral are experimenting with a garlic-based food supplement designed to alter the bovine digestive makeup to reduce the amount of methane they emit. Studies have shown the product to reduce methane emissions by about 20 percent in meat cattle, according to The New York Times.

In order to adhere to the Paris climate agreement, Mootrals owner, Thomas Hafner, believes demand will grow as governments require methane reductions from their livestock producers. We are working from the assumption that down the line every cow will be regulated to be on a methane reducer, he told The New York Times.

Meanwhile, a farm science research institute in New Zealand, AgResearch, hopes to target methane production at its source by eliminating methanogens, the microbes thought to be responsible for producing the greenhouse gas in ruminants. The AgResearch team is attempting to develop a vaccine to alter the cattle guts microbial composition, according to the BBC.

Genomic testing may also allow cattle producers to see what genes calves carry before theyre born, according to Mateescu, enabling producers to make smarter breeding decisions and select for the most desirable traits, whether it be heat tolerance, disease resistance, or carcass weight.

Despite all these efforts, questions remain as to whether biotech can ever dramatically reduce the industrys emissions or afford humane treatment to captive animals in resource-intensive operations. To many of the industrys critics, including environmental and animal rights activists, the very nature of the practice of rearing livestock for human consumption erodes the noble goal of sustainable food production. Rather than revamp the industry, these critics suggest alternatives such as meat-free diets to fulfill our need for protein. Indeed, data suggests many young consumers are already incorporating plant-based meats into their meals.

Ultimately, though, climate change may be the most pressing issue facing the cattle industry, according to Telugu of the University of Maryland, which received a grant from the Bill and Melinda Gates Foundation to improve productivity and adaptability in African cattle. We cannot breed our way out of this, he says.

Dyllan Furness is a Florida-based science and technology journalist. His work has appeared in Quartz, OneZero, and PBS, among other outlets.

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Biotechnology Could Change the Cattle Industry. Will it Succeed? - Undark Magazine

Global Genetic Engineering Vector Market Analysis Recent Trends And Regional Growth Forecast By Types And Applications 2020 – Bulletin Line

Global Genetic Engineering Vector Market Research Report Covers the existing situation and the development predictions of the industry for 2020. This Report has been prepared primarily based mostly on particular market assessment with inputs from industry experts. This estimated report consists of all have observed element about marketplace evaluation, increase Demand and forecast analysis in all over the world. This record gives a few edged examine and solution within the complicated international of polymer-based totally thermal interface materials market.

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1. Industry Overview of Genetic Engineering Vector2. Global Genetic Engineering Vector Competition Analysis by Players3. Company Profiles4. Global Genetic Engineering Vector Market Size by Type and Application (2020-2025)5. United States Genetic Engineering Vector Development Status and Outlook6. EU Genetic Engineering VectorDevelopment Status and Outlook7. Japan Genetic Engineering Vector Development Status and Outlook8. China Genetic Engineering Vector Development Status and Outlook9. India Genetic Engineering Vector Development Status and Outlook10. Southeast Asia Genetic Engineering Vector Development Status and Outlook11. Market Forecast by Regions, Type, and Application (2020-2025)12. Genetic Engineering Vector Market Dynamics13. Market Effect Factors Analysis14. Research Finding/Conclusion15. Appendix

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Allow field trials of Bt brinjal to ensure safety of crops, agri-tech body writes to govt – ThePrint

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New Delhi: The Alliance for Agri Innovation (AAI), a leading agri-tech industry body, has written to the central government and various states to allow field trials of Bt brinjal, a genetically-modified (GM) crop that was banned in 2010 following concerns raised about public health and biodiversity.

In a statement released Friday, the AAI said field trial was the only way to ensure safety of crops. blocking such trials amounts to blocking national development, it said.

AAIs letter has been sent to the agriculture ministry and chief ministers of Madhya Pradesh, Karnataka, Bihar, Chhattisgarh, Jharkhand, Tamil Nadu, Odisha and West Bengal.

The AAI statement also mentions how a 2019 peer-reviewed study had found that net returns to farmers from Bt brinjal crop averaged $2151/ha (Rs 1.61 lakh/ha) compared to $357/ha (Rs 26,768/ha) from non-Bt brinjal crop.

In India, Bt brinjal was cleared for commercial cultivation by the Genetic Engineering Appraisal Committee (GEAC) in 2009 but it was placed under an indefinite moratorium the following year by then minister of state for environment, Jairam Ramesh.

Several farmers from various states, including Maharashtra and Haryana, were found to be openly flouting the GM crop ban last year and sow Bt brinjal.

The crop has been commercially grown in Bangladesh since 2014 the first South Asian country to do so.

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Also read: Why farmers are still having to protest for their right to sow GM seeds, even in a pandemic

Brinjal is an extremely pest-prone crop, and highly susceptible to the fruit and shoot borer (FSB) pest.

To reduce loss of yield, farmers often end up spraying this crop multiple times. A report from Bangladesh had earlier noted that protein in Bt brinjaldisrupts the digestive systems of certain pests, causing them to die within three days of ingestion.

It had also stated that advantages of using Bt brinjal over non-Bt brinjal were lower pest infection, higher yields, 56 per cent reduction in environmental toxicity and fewer complaints about symptoms consistent with pesticide exposure.

Ram Kaundinya, director general of AAI, said in the statement, The deadly FSB is a menace for farmers and its caterpillar also finds its way into our homes through infected brinjals. By controlling this with Bt technology, we can save farmers income, reduce pesticide load on the environment and provide pesticide- and insect-free brinjals to consumers.

AAI head Paresh Verma added, Development of indigenous technology is in line with the Atmanirbhar Bharat mission launched by Prime Minister Narendra Modi. Delays due to a paralysis in the decision-making process by the central and state governments have essentially stalled the progress in research to develop new technologies.

The AAI statement also noted how several studies and analyses into GM cultivation over the past 23 years have found no evidence of adverse health effects on humans or animals.

Also read: Now, Bangladesh set to steal march over India with GM rice that fights malnutrition

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Allow field trials of Bt brinjal to ensure safety of crops, agri-tech body writes to govt - ThePrint

Discovery Could Lead to More Potent Garlic, Boosting Flavor and Aroma – SciTechDaily

Hannah Valentino, left, and Pablo Sobrado, right, uncover a new step in the process that makes garlic potent. Credit: Virginia Tech

For centuries, people around the world have used garlic as a spice, natural remedy, and pest deterrent but they didnt know how powerful or pungent the heads of garlic were until they tasted them.

This information changes the whole story about how garlic could be improved. Hannah Valentino, a Ph.D. candidate in the Virginia Tech College of Agriculture and Life Sciences

But what if farmers were able to grow garlic and know exactly how potent it would be? What if buyers could pick their garlic based on its might?

A team of Virginia Tech researchers recently discovered a new step in the metabolic process that produces the enzyme allicin, which leads to garlics delectable flavor and aroma, a finding that upends decades of previous scientific belief. Their work could boost the malodorous yet delicious characteristics that garlic-lovers the world over savor.

This information changes the whole story about how garlic could be improved or we could make the compounds responsible for its unique flavor, said Hannah Valentino, a College of Agriculture and Life Sciences Ph.D. candidate. This could lead to a new strain of garlic that would produce more flavor.

The discovery of this pathway opens the door for better control of production and more consistent crops, which would help farmers. Garlic could be sold as strong or weak, depending on consumer preferences.

The research was recently published in the Journal of Biological Chemistry.

Hannah Valentino, left, and Pablo Sobrado, right, are conducting research that is laying the foundation for a future in which buyers can choose garlic based on its strength and flavor profile. Credit: Virginia Tech

When Valentino, an Institute for Critical Technology and Applied Science doctoral fellow, and her team set out to test the generally accepted biological process that creates allicin, they found it just didnt happen.

Thats when the team of researchers set out to discover what was really happening in garlic.

As they peeled back the layers, they realized there was no fuel to power the previously accepted biological process that creates allicin.

By using rational design, Hannah found a potential substrate, said Pablo Sobrado, professor of biochemistry in the College of Agriculture and Life Sciences and a member of the research team. This is significant because by finding the metabolic pathway and understanding how the enzyme actually works and its structure gives us a blueprint of how allicin is created during biosynthesis.

Valentino and the team which included undergraduate students worked in the Sobrado Lab in the Fralin Life Sciences Institute directly with the substrates that comprise garlic, doing their work solely in vitro.

The researchers found that allicin, the component that gives garlic its smell and flavor, was produced by an entirely different biosynthetic process. Allyl-mercaptan reacts with flavin-containing monooxygenase, which then becomes allyl-sulfenic acid.

Importantly, the allicin levels can be tested, allowing farmers to know the strength of their crops without the need for genetic engineering. Greater flavor can simply be predicted, meaning powerful garlic could simply be bred or engineered.

We have a basic understanding of the biosynthesis of allicin that it is involved in flavor and smell, but we also now understand an enzyme that we can try to modulate, or a modify, to increase or decrease the level of the flavor molecules based on these biological processes, Sobrado said.

Because of their work, the future awaits for fields of garlic harsh enough to keep even the most terrifying vampires at bay.

There is a video with more information on this research.

Reference: Structure and function of a flavin-dependent S-monooxygenase from garlic (Allium sativum) by Hannah Valentino, Ashley C. Campbell, Jonathan P. Schuermann, Nazneen Sultana, Han G Nam, Sophie LeBlanc, John J. Tanner and Pablo Sobrado, 11 June 2020, Journal of Biological Chemistry.DOI: 10.1074/jbc.RA120.014484

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Discovery Could Lead to More Potent Garlic, Boosting Flavor and Aroma - SciTechDaily

What to Watch When Twist Bioscience Reports Fiscal Q3 Results – Motley Fool

Uncertainty is one of the central themes of investing in 2020, but investors can be certain of one thing: Valuing shares of Twist Bioscience (NASDAQ:TWST) at more than 30 times sales is a little bit ridiculous.

The company's technology platform is intriguing, but it hasn't shown signs of becoming a commercial success from a business standpoint. In fact, Twist Bioscience has reported growing losses as it has scaled its business and relied on equity financing to fund operations. That hasn't stopped the small-cap stock from soaring by more than 200% since the beginning of 2020.

On the one hand, Twist Bioscience is uniquely positioned to help customers respond to the coronavirus pandemic. It has also signed a flurry of drug discovery deals this year. On the other hand, the financial impacts of those catalysts are a bit fuzzy. Here's what metrics investors will be watching when the company reports fiscal third-quarter 2020 operating results.

Image source: Getty Images.

Twist Bioscience is developing a DNA synthesis technology platform that can be used in drug discovery, genetic engineering experiments, and diagnostic panels. It's not necessarily the technological endgame, but it's the best we have right now. Heck, the company supplies many of its competitors with synthetic DNA.

And its technology platform looks to be playing an important role in the global response to the coronavirus pandemic. Twist Bioscience has created synthetic controls of the SARS-CoV-2 virus that can be used in diagnostic tests to detect whether a person is currently infected, launched SARS-CoV-2 antibody panels for research activities, and inked drug discovery deals with biopharmaceutical companies developing therapeutic antibodies to treat COVID-19.

Additionally, the company doesn't appear to have slowed future-oriented investments in its technology platform and businesses. Twist Bioscience has signed non-COVID-19 drug discovery deals with Takeda Pharmaceuticals, Seismic Bio, and Invetx (an animal health start-up) since the end of the fiscal second quarter of 2020.

The biggest question facing investors is pretty simple: Will announcements translate into tangible progress?

Investors are assuming Twist Bioscience will report a noticeable bump in quarterly revenue from coronavirus-oriented research tools, although financial details weren't disclosed for any of the drug discovery deals. The latter appear to be dependent on technology access fees and milestone or royalty payments, which wouldn't necessarily have a significant near-term impact.

Here's the problem: Even an incredible quarter probably isn't enough to make sense of the stock's current valuation. Investors shouldn't be paying 30 times sales for a money-losing business that has grown operating losses faster than gross profit and is entirely reliant on stock offerings to keep the lights on.

In other words, even a relatively strong quarter could cause the stock to drop. There's simply too much success baked into the company's market valuation right now. It's difficult to see how Twist Bioscience's $3 billion valuation is sustainable with publicly available information. Then again, it's 2020. Many stock valuations don't make much sense right now.

A great business is always a great investment in the long run, but a great investment is not always a great business. Thus far in 2020, Twist Bioscience has been a great investment, but the stock's ascension is detached from business fundamentals. That's likely to be true even with an amazing increase in revenue from the coronavirus pandemic, assuming that's what the results show. Investors might want to take that into account, as starting or adding to a position at current prices might sap long-term returns. Unless the company announces a major event that's not currently on investors' radar, it's simply difficult to see shares holding onto recent gains.

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What to Watch When Twist Bioscience Reports Fiscal Q3 Results - Motley Fool

GentiBio Joins Cell Therapy Chase With $20M and New Treg Technology – Xconomy

XconomyBoston

One of the challenges facing cell therapy developers is collecting enough cells to produce a viable treatment. Its a particularly pronounced problem for therapies employing regulatory T cells (Tregs), a type of immune cell thats relatively scarce in the blood, says GentiBio CEO Adel Nada.

Some biotech companies are developing Treg cell therapies from a patients own Tregs. GentiBio makes its Treg therapeutic candidates from an entirely different type of immune cell, and Nada says this approach could make Treg cell therapy production more scalable. On Wednesday, the Boston-based startup announced its launch backed by $20 million in funding.

The immune system is comprised of many different types of cells that have different functions. For example, T cells seek out and destroy pathogens, and they also prompt other immune cells to mount a response. Tregs do the opposite, regulating or suppressing an excessive immune response. Such overreactions are associated with some autoimmune disorders, making Treg cell therapies promising as a way to treat them.

Rather than harvesting a patients Tregs, engineering those cells, and then multiplying them in a lab, GentiBio works with immune cells called CD4+, which are also known as helper T cells. In addition to playing multiple roles in an immune response, these cells are abundant in the blood. GentiBio uses genetic engineering techniques to make Treg-like cells from CD4+ cells. Scientists have already shown that this approach can work in animals. Not only has the technology produced these engineered Tregs, but these cells have also shown the potential to address graft-versus-host disease and encephalitis in mice. Results were published in June in the journal Science Translational Medicine.

GentiBios Tregs are engineered with additional features. Attaching a T-cell receptor to these cells enables them to target specific tissues, Nada says. He adds that synthetic biology technology helps these cells survive longer than typical Tregs. Once infused into a patient, these cells would be tunable, meaning that their numbers could be dialed up or down to the level needed to treat a particular disease. Nada declined to disclose his companys disease targets, other than to say that they are autoimmune diseases of high unmet medical need.

A growing number of companies are researching Treg therapies as a way to treat various diseases. Sonoma Biosciences launched in February, revealing $40 million in Series A financing and its plans to engineer cell therapies from a patients own Tregs. The startup, which splits its operations between South San Francisco and Seattle, has not disclosed its disease targets but co-founder and CEO Jeffrey Bluestone told Xconomy that in addition to autoimmune disorders, the companys approach has potential applications in treating cancer and neurodegeneration.

Pandion Therapeutics (NASDAQ: PAND) is developing drugs intended to treat disease by multiplying Tregs throughout the body without activating inflammatory cells. Last month, the Watertown, MA-based biotechs IPO raised $135 million, part of which will support PT101, the companys lead drug candidate that is currently in early-stage testing in moderate-to-severe ulcerative colitis. San Francisco-based Orca Biosciences is developing proprietary mixtures of various types of immune cells, including Tregs, to address disease. One of its programs, a combination of T cells and Tregs, is currently in Phase 1/2 testing in blood cancers.

GentiBios approach can be used to make Treg therapies from a patients own cells, as well as off-the-shelf therapies produced from the cells of healthy donors. Nada says its too early to talk about which type the company is developing. The companys research is based on technologies licensed from Seattle Childrens Hospital and Research Institute, the Virginia Mason Health System-affiliated Benaroya Research Institute in Seattle, and Israels MIGAL Galilee Research Institute.

With the academic collaborators weve been working with, weve generated preclinical data that can support regulatory filings [for clinical trials], Nada says. We have assets that allow us to steadfastly march to the clinic, at a pace that is not what you would expect from an academically grown asset.

GentiBios financing, a seed round, was led by OrbiMed, Novartis Venture Fund, and RA Capital Management. Nada says the new cash enables his company to build infrastructure, including manufacturing, to support early-phase clinical testing in two indications, as well as non-clinical research for its other programs. GentiBio will need to raise more money next year to support those additional programs, Nada says.

Photo by Flickr user Alachua County via a Creative Commons license

Frank Vinluan is an Xconomy editor based in Research Triangle Park. You can reach him at fvinluan@xconomy.com.

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GentiBio Joins Cell Therapy Chase With $20M and New Treg Technology - Xconomy

Viewpoint: Battling deadly disease with gene drives is worth the limited risk – Genetic Literacy Project

The fate of society rests in part on how humans navigate their complicated relationship with insects trying to save good insects and control bad ones. Some insects, like mosquitoes, bite people and make them sick remember Zika? Now the U.S. mosquito season is already in full swing, withover 10 cases of Dengue feverreported in the Florida Keys this year. Some insects, likebees, are pollinatorsthat help produce our food. Others, likelocusts, currently threaten cropsin East AfricaandAsia, preferring to eat our food instead.

Insects have proven themselves extremely capable at evolving strategies to get around control methods, such as chemical insecticides and habitat modification, and current pest control technologies are simply not keeping up.

We arebothinsect scientists. Our research has includedengineering a fungus to control malaria mosquitoes,uncovering the reproductive biology of honey bee workersand understanding thehealth impacts of invasive ticks. Weve come to appreciate the potential of emerging technologies like gene drive. This technology can guarantee that a trait will be inherited by the next generation. Such traits include making mosquitoesimmune to the malaria parasiteso they cannot spread the disease to humans.

Recently we contributed to astatement that advocates for continuing gene drive research. In light ofcalls for a moratorium, this statement recognizes that a ban on gene drive research would hamper a better understanding, and thus mitigation, of risks associated with this technology.

Moratoriums on gene drive technology have been called for and rejectedat the last twoUnited Nations Conventions on Biological Diversity. But there isa new push for a moratorium.

Gene drive is a technology that could allow society to control insects in a more targeted manner.

The general underlying principle of all gene drives is an organism that will produce offspring similar to themselves.

Some characteristics are randomly passed on from parents to the next generation. However, gene drive forces a different type of inheritance that ensures a specific characteristic is always present in the next generation. Scientists engineer gene drive using various molecular tools.

Gene drive is not just a human invention; some occur naturally in insects. For example, instalk-eyed flies, a gene on a sex-related chromosome causes any male fly to die without a certain gene cargo, including a gene that results in longer eyestalks. This type of genetic phenomenon has been well studied by scientists.

To date, gene drive has been discussed in the media primarilyin order to eradicate malaria. This may give you the impression that gene drive can be used onlyto drive mosquitoes to extinction. However, gene drive technologies are highly versatile and can be designed to bring about different outcomes. They can also be applied in most insect species that scientists can study in the laboratory.

Insects reproduce quickly and produce lots of offspring, which makes them obvious candidates for a technology that relies on inheritance like gene drive. This is why insects are at the leading edge of gene drive research. Gene drive is a new technology that could provide a solution to a variety of insect issues society faces today.

For instance, a gene drive has beendeveloped to stop a major crop pest, the spotted-wing Drosophila. Insecticide sensitivity could be spread through populations of this pest species to stoptens of millions of dollars in crop damage every yearin the United States.

Gene drive could also be a more targeted approach to stopping invasive insects, such as the infamousfire ant, from destroying native ecosystems. In the United States, millions of dollars have been spent onremoving fire ants using techniques including chemical insecticides, but if these persistent ants are not completely eradicated, they invade again.

Aside from how good insects are at circumventing our strategies to control them, another major struggle for controlling insects is finding them. Insects have evolved to quickly find the opposite sex to mate, and gene drives, which are passed on by mating, can take advantage of this fact of insect life. This also means this technology targets only the intended species, which is not the case for chemical insecticides currently in use.

Insect scientists, inspired by natural examples of gene drive, have wanted to design gene drive in insects for decades. Only recently have new molecular tools, such as the gene editing toolCRISPR-Cas, made the gene drive dream a reality. For now, gene drive insectslive in laboratories and none has been released into the wild. Still, a lot can be learned about how gene drive works while it is safely contained in a laboratory.

Using gene drive is not a universally popular idea. Criticisms tend to fall into three categories: ethical concerns, mistrust of technology and unintended ecological consequences.

Ethical concerns about gene drive are often motivated by larger issues, such as how to stop gene drive from being used in biological weapons by engineering insects that are more dangerous. Then there is the question of who should decide which gene drive projects move forward and what types of insects with gene drive can be released into the environment. These questions cant be answered by scientists alone.

Societal mistrust of technologyis a hurdle that some powerful, innovative technologies must overcome for public acceptance. The issue of technological mistrust often stems from disagreements about who should be developing technology to control insects and for what purposes.

The third common argument against gene drive technologies is that they might cause unintended consequences in the ecosystem because gene drive is designed by humans and unnatural. What will happen to the natural ecosystem if a population, even of mosquitoes that make people sick, is driven to extinction? Will this cause threats to natural biodiversity and the security of food? These questions are ultimately asking the consequences of intervening in the natural order of the world. But who defines what is the natural state of an ecosystem?Ecosystems are already constantly in flux.

When a gene drive is developed, it is tailored to the needs of a particular situation. This means theanticipated risksposed by each gene drive are project-specific and should be considered and regulated on a case-by-case basis. A responsible way to protect society from these risks is to advocate for continued research that enables scientists to describe and find solutions to them. Beyond the science, regulatory and accountability systems are needed so that regulations are adhered to and public safety is protected.

Researchers are also still exploring the science underlying the gene drive. Can gene drive be designed to be reversible or more efficient? Can the effect of a gene drive on an ecosystem be predicted? Such important unanswered questions are why even the most ardent supporters of this technology say more research is needed. Society needs new tools to control insect pests and protect ecosystems, and gene drive promises to augment our toolbox.

Isobel Ronai is an Endeavour Postdoctoral Research Fellow at Columbia University. Follow her on Twitter @IsobelRonai

Brian Lovett is a postdoctoral researcher at the Division of Plant and Soil Sciences at West Virginia University working on fungal biology and biotechnology. Follow him on Twitter @lovettbr

This article was originally published at the Conversation and has been republished here with permission. Follow the Conversation on Twitter @ConversationUS

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Viewpoint: Battling deadly disease with gene drives is worth the limited risk - Genetic Literacy Project


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