Monthly Archives: June 2021

Global Genome Editing/Genome Engineering Market is valued approximately USD 4.4 billion in 2019 and is anticipated to grow with a healthy growth rate of more than 17.00 % over the forecast period 2020-2027.

Genome Engineering technique is used for deletion, insertion and modification of genome of a microorganism. Genome editing/genome engineering plays an integral role in modern-day biology and is widely used in the biopharmaceutical and biotechnology industry to alter the genome of microorganism to perform process such as fermentation which yield desired products. Also, this approach is majorly used for understanding DNA in cells of organism to have a better understanding of their biology, to treat infectious and autoimmune diseases. Availability of government funding and growth in the number of genomics projects are the few factors responsible for growth of the market over the forecast period of 2020-2027. For Instance: in 2017, in Canada, as per the University of Guelph, University of Genome Canada Bioinformatics and Computational Biology along with other eligible sources provide funding of $12 million for different genomics-based research projects. Similarly, according the Japan Agency for Medical Research and Development Organization, in 2017, government of Japan has introduced various initiatives such as Tohoku Medical Megabank project, Platform Program for promotion of Genome Medicine etc., these projects are inclined to provide research infrastructure.

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Such government initiatives would increase the need for genome editing and genome engineering. Also, the key players of global Genome Editing/Genome Engineering market have adopted various strategies to gain competitive advantage including product launch, innovation, technological advancements, investment, funding and others. However, high equipment cost is the major factor restraining the growth of global Genome Editing/Genome Engineering market during the forecast period.

The regional analysis of global Genome Editing/Genome Engineering market is considered for the key regions such as Asia Pacific, North America, Europe, Latin America and Rest of the World. North America is the leading/significant region across the world in terms of market share as the region is one of the most significant markets for development of gene therapy in the US, the the rising prevalence of infectious diseases and cancer, increasing use of genetically modified crops, and the availability of research grants and funding. Whereas Asia-Pacific is also anticipated to exhibit highest growth rate / CAGR over the forecast period 2020-2027.

Major market player included in this report are:Thermo Fisher ScientificMerckHorizon DiscoveryGenscriptSangamo TherapeuticsLonzaEditas MedicineCrispr TherapeuticsEurofins ScientificPrecision Biosciences

The objective of the study is to define market sizes of different segments & countries in recent years and to forecast the values to the coming eight years. The report is designed to incorporate both qualitative and quantitative aspects of the industry within each of the regions and countries involved in the study. Furthermore, the report also caters the detailed information about the crucial aspects such as driving factors & challenges which will define the future growth of the market. Additionally, the report shall also incorporate available opportunities in micro markets for stakeholders to invest along with the detailed analysis of competitive landscape and product offerings of key players. The detailed segments and sub-segment of the market are explained below:

By Technology:CRISPRTALENZFNANTISENSEOther technologiesBy Product and Services:Reagents and ConsumablesSoftware & systemsServicesBy Application:Cell line EngineeringGenetic EngineeringDiagnostics ApplicationsDrug discovery and developmentOthersBy End-user:Pharmaceuticals CompaniesBiotechnology CompaniesAcademic and Government research institutesBy Region:North AmericaU.S.CanadaEuropeUKGermanyFranceSpainItalyROE

Asia PacificChinaIndiaJapanAustraliaSouth KoreaRoAPACLatin AmericaBrazilMexicoRest of the World

Furthermore, years considered for the study are as follows:

Historical year 2017, 2018Base year 2019Forecast period 2020 to 2027

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Target Audience of the Global Genome Editing/Genome Engineering Market in Market Study:

Key Consulting Companies & AdvisorsLarge, medium-sized, and small enterprisesVenture capitalistsValue-Added Resellers (VARs)Third-party knowledge providersInvestment bankersInvestors

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Genome Editing/Genome Engineering Market expectation surges with rising demand and changing trends by industry analysis through 2027 The Manomet...

The World Health Organisation maintains the coronavirus most likely arose in bats, and then spread to humans via an as-yet unidentified intermediary animal. Image / CDC

Despite the "lab leak theory" taking flight again in the US media, global scientists still point out there's little evidence to suggest the SARS-CoV-2 virus originated in a Wuhan laboratory and a large amount to suggest it came from nature. Otago University virologist Dr Jemma Geoghegan and Massey University infectious disease ecologist Professor David Hayman set out three reasons why a lab-made pandemic remains extremely unlikely.

There's a strong precedent for coronaviruses that have become "zoonotic", or jumped from animals to humans.

There have been seven that scientists are aware of including SARS-CoV and MERS-CoV and bats are thought to have been involved in most of these.

"And five of these human coronaviruses have emerged in the last 20 years," Geoghegan said.

While the source of the SARS-CoV-2 virus still hasn't been pinned down, she said that wasn't unusual.

"In fact, we don't know where most of the viruses that infect us have come from," she said.

"This is why we need to sample more viruses in nature and expand our knowledge of the diversity of viruses that exist."

Hayman added that, now that scientists were looking even harder, more cases of infections from newly-detected or "novel" coronaviruses were coming to light.

"One was identified in pneumonia patients in Malaysia where people were living in villages in close contact with domestic and wild animals."

While the pandemic found the world poorly prepared for it, scientists had been warning for years that the growing interaction between animals and nature particularly through habitat destruction had been raising the risk of a catastrophe like Covid-19.

Just like the first Sars coronavirus, the first cases of Sars-Cov-2 were associated with an animal market this time in Wuhan, China.

The World Health Organisation's report identified that live animals like ferret-badgers and rabbits were being traded in these markets.

"These animals could provide an intermediate host for the virus to jump to humans," Geoghegan said.

"It's exactly the type of place you'd expect a zoonosis event to happen."

Hayman added that some of the farmed species had complex commodity chains.

That meant the farms could well be in places where there was a greater diversity of bat viruses than in Wuhan, where the pandemic appeared to have started.

"And we have seen how SARS-CoV-2 can be maintained in farmed fur animals, such as the very large outbreaks in farmed mink in Europe, which led to substantial mink to mink transmission as well as mink-to-human transmission."

Other evidence also shows that this type of coronavirus has existed in bats for decades and the SARS-CoV-2 genome sequence happened to be 96 per cent identical to a coronavirus found in horseshoe bats.

In one of the earliest major studies into the virus, scientists analysed its genetic template for spike proteins or armatures on the outside of the virus that it used to grab and penetrate the outer walls of human and animal cells.

More specifically, they focused on two important features of the spike protein.

Those were its receptor-binding domain (RBD) - a kind of grappling hook that grips on to host cells - and what's called the cleavage site, a molecular can opener that allows the virus to crack open and enter host cells.

They found the RBD portion of the SARS-CoV-2 spike proteins had evolved to effectively target a molecular feature on the outside of human cells called ACE2 - a receptor involved in regulating blood pressure.

The SARS-CoV-2 spike protein was so effective at binding the human cells, in fact, that the scientists concluded it was the result of natural selection, and not the product of genetic engineering that some theorists have suspected.

The idea of natural evolution was given further credence by data on the virus' backbone - its overall molecular structure.

If someone were seeking to engineer a new coronavirus as a pathogen, they would have constructed it from the backbone of a virus known to cause illness.

But the scientists found that the backbone differed greatly from those of already known coronaviruses.

It turned out to mostly resemble related viruses found in bats and pangolins - scaly-skinned mammals that are prized delicacies in China.

That led scientists to suspect one of two possible scenarios.

In one scenario, the virus evolved to its current state through natural selection in an animal host and then jumped to humans.

Yet there were no documented cases of direct bat-human transmission, suggesting that an intermediate host was likely involved between bats and humans.

In this scenario, both of the distinctive features of SARS-CoV-2's spike protein - the RBD portion that binds to cells and the cleavage site that opens the virus up - would have evolved to their current state before entering humans.

In this case, the current epidemic would probably have emerged rapidly as soon as humans were infected, as the virus would have already evolved the features that made it pathogenic, or able to spread between people.

In the other proposed scenario, a non-pathogenic version of the virus jumped from an animal host into humans and then evolved to its current state within the human population.

A coronavirus from a pangolin could possibly have been transmitted to a human, either directly or through an intermediary host such as civets or ferrets.

After that, the other distinct spike protein characteristic of SARS-CoV-2 - the cleavage site - could have evolved within a human host, or possibly among a group of people, before the outbreak kicked off.

Geoghegan said the idea that the cleavage site was so unusual that it must have been engineered was "totally false".

She said that assumed an amino acid sequence within the site called PRRAR had been created in a lab.

Yet these cleavage sites had been found in other coronaviruses - even with the exact same "PRRAR" insert.

"It's a totally bonkers argument," Hayman added.

"Similarly, people really need to understand that these viruses do recombine. For example, the novel virus from Malaysia that was recently detected seems to be a recombinant of a cat and dog viruses, which were also previously not known."

The slightly more plausible alternative lab leak theory was that scientists could simply have been growing a culture of the virus, and it escaped from there.

But that would've had to assume the virus could have leaked from a secure research facility and also neglected the fact that the virus' feature of entry and infection markedly diminished in a lab culture setting.

And then, as Dr Jonathan Stoye, group leader of the Retrovirus-Host Interactions Laboratory at the UK's Francis Crick Institute, pointed out, the virus' spread around the world didn't gel with the lab-grown theory.

"The genome of SARS-CoV-2 shows more than 1000 individual differences from its closest known relative," he said.

"Given the rate of nucleotide change observed in virus spreading through the human population over the past year it seems extremely improbable, perhaps impossible, that changes spanning such an evolutionary distance could have occurred during virus growth in a lab.

"It therefore remains most likely that the immediate ancestor to SARS-CoV-2 exists in the wild and is still to be found."

Scientists aren't saying the possibility of a lab leak should be entirely ruled out on the contrary, many argue that it should be comprehensively investigated.

But more than a year after the outbreak, the weight of evidence overwhelmingly points to a natural source, while there's little to suggest the virus came from a lab.

Part of the lab leak theory is predicated on the fact that the Wuhan Institute of Virology has carried out extensive work on coronaviruses in bats.

While the institute didn't shared its lab records with a team of WHO investigators, there's as yet been no evidence that any samples of the virus were kept there before it was first reported, nor were there any viruses that could have combined to create it.

Much of the recent coverage has been fueled by a US intelligence report that stated several researchers had become sick with "symptoms consistent with both Covid-19 and common seasonal illness".

A top director at the institute has rejected this inference, reporting that all staff have tested negative for Covid-19 antibodies, and that there'd been no turnover of staff in the coronavirus team.

"Why would these scientists be working on a random secret bat virus and not have published anything on it previously?" Geoghegan said.

"Those that do gain-of-function experiments work on really well characterised viruses, and there would be really close genetic relatives of the virus published before.

"But for SARS-CoV-2, there isn't. Even the closest viruses in bats and pangolins are too divergent to be a starting point."

Hayman added that every expert in the area would have asked themselves, "what if it was that lab?"

"We ask ourselves, 'What would we see, what evidence would we need to support it?', and even, 'have we been lied to?'. We have, most of us, agonised over the possibilities.

"But right now the only way that this can be true is if there is a massive conspiracy, because while the lab pathway is a potential pathway, nothing published or reliably reported by China to date supports a lab escape at all, and there is a huge amount of data to support this being a natural event.

"This discussion would have gone away if it hadn't have happened in China. But unfortunately finding conclusive evidence of either is very unlikely."

See the article here:
Covid 19 coronavirus: Why the lab leak theory is still unlikely - New Zealand Herald

Markeshaw Tiruneh G/Medhin,1 Endeshaw Chekol Abebe,2 Tekeba Sisay,3 Nega Berhane,3 Tesfahun Bekele Snr,1 Tadesse Asmamaw Dejenie1

1Department of Biochemistry, School of Medicine, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia; 2Department of Biochemistry, College of Health Sciences, Debre Tabor University, Debre Tabor, Ethiopia; 3Institute of Biotechnology, College of Natural Science, University of Gondar, Gondar, Ethiopia

Correspondence: Markeshaw Tiruneh G/Medhin Tel +251922712112Email [emailprotected]

Abstract: Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated proteins are referred to as CRISPR-Cas9. Bacteria and archaea have an adaptive (acquired) immune system. As a result, developing the best single regulated RNA and Cas9 endonuclease proteins and implementing the method in clinical practice would aid in the treatment of diseases of various origins, including lung cancers. This seminar aims to provide an overview of CRISPR-Cas9 technology, as well as current and potential applications and perspectives for the method, as well as its mechanism of action in lung cancer therapy. This technology can be used to treat lung cancer in two different ways. The first approach involves creating single directed RNA and Cas9 proteins and then distributing them to cancer cells using suitable methods. Single directed RNA looks directly at the lungs mutated epidermal growth factor receptor and makes a complementary match, which is then cleaved with Cas9 protein, slowing cancer progression. The second method is to manipulate the expression of ligand-receptors on immune lymphocytic cells. For example, if the CRISPR-Cas9 system disables the expression of cancer receptors on lymphocytes, it decreases the contact between the tumor cell and its ligand-receptor, thus slowing cancer progression.

Keywords: CRISPR, Cas9, CRISPR-Cas9 technology, cancer, lung cancer, cancer treatment

The word CRISPR-Cas9 refers to Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated proteins.15 CRISPR-Cas9 system is a kind of acquired immunity possessed by most bacteria and archaea (prokaryotes) to act against their enemies (bacteriophages).4,6 It is a ribonucleic acid (RNA) guided, convenient, and versatile endonuclease platform for site-specific genome editing,1,7,8 which can play a tremendous role in the application of cancer therapy.1 The application of this technology can be used to resolve mutations and to introduce site-specific therapeutic genes in human cells so that, correcting disease-causing mutations, and alleviate disease-related symptoms. This system is also a useful tool for delineating molecular mechanisms involving hematological malignancies.4 Sequence-specific gene editing using CRISPR-Cas9 shows promise as a novel therapeutic approach for the treatment of a variety of diseases that currently lack effective treatments like cancers.3,9 To accomplish its task, it requires Cas9 DNA endonuclease protein and single guided RNA (sgRNA) that can produce precise gene matching for editing and correction techniques.2 So the system has enabled easy manipulation of genes for the scientific community by making the hybrid to the target sequence and cleaving the double-strand DNA.10

Additionally, the CRISPR-Cas9 technology is increasingly feasible to overcome drug resistance in breast cancer therapy and will become an essential tool for personalized medicine.4 It is a technological breakthrough that facilitates the ability to change nucleic acids,11 and with continued improvement in the function, the system can help to develop best treatment options to a variety of genetic disease which affects several tissues in our body.12 Gene manipulation using CRISPR-Cas9 system has revolutionized and made it easy to study the work of genes and importantly opens the new era of treatment mechanisms for different disease conditions including cancer.13 Technologies like this are a simple and efficient method of targeting the required DNA regions.14 Thus, scientists have designed two main components of the system for easy detection and alteration of gene function one component is a protein Cas9 that enzymatically cleave the desired gene and the sgRNA which scans and determines where the gene of interest will be cleaved by Cas9 protein.3,12,15 The system has been scientifically optimized and developed to regulate expression of the gene, modify and edit the desired locus and this makes the technology of choice seen by the scientific community to treat or edit disease-causing mutations more efficiently than ever before. Furthermore, its application is encouraging for more vigorous gene therapy in clinical setups.16 Based on the discovery, there are three main types of CRISPR-Cas9 system (I to III) and with three additional types (IV to VI) being identified more recently.17 They are different during the processes of immunity, adaptation, expression, and interference, each type acts in distinct mechanisms to ensure genetic manipulation. Type I employs a large complex of Cas9 proteins with distinct helicase and DNase activities, while type III employs repeat-associated mysterious proteins, which form a large Cas9 superfamily. Another classification is based on subunit effectors, with multi-subunit effector complexes being the most common. Types I, III, and IV are grouped in class 1. Those systems, on the other hand, that have a single subunit effector are categorized as class 2 comprising types II, V, and VI.17,18 Type II uses only a Single protein (Cas9) for its nuclease activity and has got more attention and adopted for genome engineering.5,17,19 Thus, the objective of this seminar is to introduce CRISPR-Cas9 technology and describe current applications and future perspectives of the system with its mechanism of action on lung cancer therapy.

The tracrRNA gene will be transcribed to tracrRNA, the crRNA gene will be transcribed to pre crRNA, and the Cas9 gene will be transcribed to Cas9 messenger RNA and converted to Cas9 protein, all of which will be post-transcribed and chopped off to form the mature CRISPR-Cas9 complex.20 Cas1 and Cas2 integrase, which are present in all CRISPR forms, catalyze spacer integration on the CRISPR array especially on the leader end of the repeat there will be a nucleophilic attack of the 3 OH of the protospacers followed by the same practice on the spacer end of the repeat.21

The CRISPR-Cas9 method has a variety of formulation methods for genome editing. The use of a plasmid-based CRISPR-Cas9 system encoding both the Cas9 protein and sgRNA from the same vector, which is necessary to avoid multiple transfections of different components of the technology, is the leading and possibly the easiest technique. The Cas9 protein and sgRNA will be expressed in the vector, which will form the sgRNA-Cas9 complex within cells to edit the target genomic sequences.3,12,15,18 The second approach involves combining Cas9 mRNA and sgRNA. The sgRNA-Cas9 complex will be formed when Cas9 mRNA is converted into Cas9 protein in cells. The third strategy is to deliver the in vitro assembled sgRNA-Cas9 complex directly to the cell.18

It is difficult to transmit nucleic acid in general, and CRISPR-Cas9 in particular, to the target tissue or cell. Physical and vector (viral or non-viral) approaches are two of the most widely used distribution strategies.11,22,23 Electroporation and microinjections are used in physical methods, while viral delivery strategies such as adeno associated virus (AAV) are widely used in vector-based methods since they are not disease-causing agents and can infect both dividing and non-dividing cells25 and lentivirus with inactivated integrase enzymes are under investigation.24

Another technique is lipofection (lipid-mediated nanoparticle transfection), which is possibly the most efficient CRISPR-Cas9 in vivo delivery method.22 This technique was further developed by26 and is currently being tested in clinical trials.13,24

The CRISPR-Cas9 system, as discussed, a little earlier, is made up of two main components that work together to accomplish its goal.19 The sgRNA contains crRNA, which scans and identifies the target DNA sequences that must be cleaved and corrected, and transactivated crRNA (tracrRNA), which recruits component two, the Cas9 protein DNA endonuclease, which can sense, identify, and establish site-specific double-strand DNA breaks (DSB).15 Because of its simplicity and convenience, the bacterial type II CRISPR-Cas9 system has been used for RNA-guided engineering nucleases.4,18 However, the proto-spacer adjacent motifs (PAM) sequences are required by the method. After recognition, two Cas9 domains cleave double-stranded DNA: the endonuclease domain named for characteristic histidine and asparagine residues (HNH) domain, which cleaves the complementary strand, and the endonuclease domain named for an E. coli protein involved in DNA repair (RuvC-like) domain, which cleaves the non-complementary strand.17 As a result, the host DNA repair machinery introduces numerous mutations such as substitutions, deletions, and insertions in the target genome, including non-homologous end joining (NHEJ) or homologous-dependent repair (HDR).1518 Another paper, CRISPR-Cas9 for Cancer Therapy: Hopes and Challenges, supports this theory by demonstrating that the sgRNA-Cas9 complex scans and anneals to the genomic target sequence with base-pairing complementarity and precisely cleaves double-stranded DNA of the target cell after identification of the protospacer adjacent motif (PAM) sequence adjacent to the target sequence. NHEJ or HDR pathways are activated as a result of double-strand breaks. NHEJ is an error-prone repair mechanism that results in indels (insertions or deletions) of random base pairs disrupting the target sequence in the absence of a homologous repair prototype with more specific repair mechanisms.23,27

Lung cancer is the major cause of death in the United States and a significant public health concern worldwide.5 In both developed and developing countries, it is a common cause of morbidity and mortality.28 According to a study conducted by the American Lung Cancer Society in 2015, lung cancer claims the lives of almost 150,000 people each year. However, surgery and radiation were used as treatment options. The treatment was later changed to selective Tyrosine kinase inhibitors (TKIs) like gefitinib and erlotinib to inhibit the tyrosine kinase activity of epidermal growth factor receptor, which has technical difficulties and nonspecific cytotoxicity (EGFR).29,30 Extracellular ligand binding, transmembrane, and intracellular tyrosine kinase domains are found in this membrane glycoprotein. When the ligand activator binds to the extracellular ligand domain, it transduces and initiates intracellular kinase activities, which cause cellular proliferation, neovascularization, invasion, and metastasis, as well as reduced apoptosis and glycolysis activation. These medications, however, have encountered drug resistance.28,29

The CRISPR-Cas9 device is the start of a new biotechnological era and a groundbreaking technology that is being used to treat lung cancer.6,29 The system works in two ways. The first is by designing sgRNA that looks for the mutated EGFR sequence, which is then accompanied by Cas9 protein. To do so, scientists created a CRISPR device that has complementary sequences with the mutated EGFR and introduced it into the patient, as mentioned earlier which has complementary sequences with the mutated EGFR and introducing this into the patient. As this complementary sequence binds to the mutated EGFR, the Cas9 protein (endonuclease) creates a double-stranded or single-stranded DNA break, depending on the type of enzyme used, followed by DNA repair mechanisms such as homologous or non-homologous DNA repair.29 If the receptor mutation is limited, there will be no contact between the ligand activator, resulting in no cell proliferation, neovascularization, or cancer metastasis, and the problem will be solved. The inhibition of EGFR by CRISPR-Cas9 increases the expression of major histocompatibility complex class I, which improves cytotoxic lymphocyte recognition and lysis of tumor cells.30,31 Off-target effects, which can induce genome instability, gene functional disturbances, and epigenetic alterations, are a challenge. Off-target effects of CRISPR-Cas9 systems, particularly when used for therapeutic purposes, should be minimized and precisely profiled. Off-target effects are separated into two categories: off-target binding and off-target cleavage. Cas9 can bind to target sequences that are partially complementary to sgRNA and inhibit target gene transcription without cleaving them.8 Off-target binding effects may thus be removed in traditional off-target identification approaches, such as using in vitro assembled sgRNA with a long-lasting association with cas9, which also has a high proportion of on-target and high efficiency for genome editing. Another technique is to use a Cas9 variant or modified Cas9 that can generate a single nick at one strand.23 So that the off-target effect is reduced.

The second, and equally significant, strategy for using this biotechnological method to treat lung cancer is to search for immune cells like lymphocytes. T cells are immune cells that are extracted from the blood of patients engaged in a clinical trial for lung cancer treatment in China, and then CRISPR-Cas9 is used to knock out a gene in the cells that encodes a protein called PD-1. The edited gene cells would then be propagated in the lab before being injected back into the patients bloodstream.6,25 Scientists took blood from the patient and extracted lymphocytes, which were then treated with a CRISPR-Cas9 gene-editing system containing a sgRNA sequence with a pattern identical to lymphocytes programmable death 1 protein (PD 1). When the system detects its target sequence, cas9 would sever the DNA, which is then repaired by cell repair mechanisms. When the expression of the PD 1 gene is blocked or disabled, cancerous cells lack the receptor on immune lymphocytes.6,25 As a result, if lymphocytes do not express the PD 1 receptor well, there will be less contact between the cancerous ligand and receptor, causing the T cell receptor to identify the problematic cell and perform its function. Naturally, these manipulated lymphocytes were screened for viability and lympho-proliferation to rule out new mutations, and only those cells that passed the test were returned to the patient.6,25 Furthermore, knocking out the PD-1 protein on immune cells is necessary for caspase activation, which is needed for programmed cell death and enhanced apoptosis in cancerous cells.31 It also concludes that PD-1-deficient cells have potent antitumor activity of cytotoxic lymphocytes. The hyperactivity of the manipulated T cells is one of the technologys drawbacks for use in this way6 and obtaining a safe and efficient delivery method, as well as some side effects Patients with advanced NSCLC with positive PD-1 expression were assigned to a Phase I clinical trial to assess the safety of CRISPR-Cas9-mediated knockout of PD-1 gene therapy in patients with metastatic non-small cell lung cancer. Nine patients were enrolled, and eight patients received PD-1 deficient T cell therapy, and the patients were manifested with PD-1 deficient T cell therapy.25 Patients undergoing PD-1 deficient T cell therapy, on the other hand, appeared to be healthy, and researchers recommended that broader studies be conducted to determine the most appropriate dosage and immune response.

In cancer biology, the CRISPR-Cas9 device has a bright future ahead of it,9, because it is a technology that is adaptable, simple, convenient and efficient.32,33 The method introduces a novel approach to cancer treatment by allowing for modifications to the genome of target cells, which was previously difficult to achieve.3436 the technologys versatility, effectiveness, and flexibility would make it the best form of cancer care in the future.4,37,38 It will affect cancer biology as a whole in the future,34 and if researchers have devised well-organized strategies and instruments for delivering the technology to the target cell or tissue, as well as effective methods and instructions for controlling and eliminating the technologys off-target effects.

The CRISPR-Cas9 device is a recent biotechnological breakthrough and scientific achievement. This technology has created a new treatment option for diseases of various origins, such as cancer and infectious disease. To solve the problem, the best sgRNA must be designed using a CRISPR tool (http://crispr.mit.edu) and its associated endonuclease cas9 protein against the target sequence. However, ethical concerns, the need for the best delivery strategies, and the risk of off-target effects are only a few of the problems that must be addressed. Since the technology is still in its infancy, researchers must devise simple methods and mechanisms to track and test its protection and efficacy. For a simple comparison, the benefits of this technology are simple, fast, relatively effective, relatively precise, and versatile, while the drawbacks are distribution is difficult, ethical problems are highly conservative, some off-target effects, and some adverse effects.

ATP, Adenosine triphosphate; CRISPR, Clustered regularly interspaced short palindromic repeat; CRISPR-Cas, Clustered regularly interspaced short palindromic repeat-associated; CrRNA, Clustered regularly interspaced short palindromic repeat ribonucleic acid; DNA, Deoxyribonucleic acid; DSB, Double-stranded break; EGFR, epidermal growth factor receptor; HDR, Homologous directed repair; mRNA, Messenger ribonucleic acid; NHEJ, Non-homologous end-joining; PD 1, Programmable death protein 1; RNA, Ribonucleic acid; SgRNA, Single guided ribonucleic acid; TracrRNA, Trans activating clustered regularly short palindromic repeat ribonucleic acid; TKIs, Tyrosine kinase inhibitors.

All authors made substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; agreed to submit to the current journal; gave final approval of the version to be published; and agree to be accountable for all aspects of the work.

There is no funding to report.

The authors declare that they have no conflicts of interest for this work.

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