February 7, 2017 A cartoon shows gene editing through engineered CRISPR\/Cas9En delivery in the Rotello lab at UMass Amherst. The researchers have overcome an obstacle in the technology by designing a delivery system using nanoparticles to assist CRISPR\/Cas9 across the cell membrane and into the nucleus while avoiding entrapment by cellular machinery. Credit: UMass Amherst <\/p>\n
More and more scientists are using the powerful new gene-editing tool known as CRISPR\/Cas9, a technology isolated from bacteria, that holds promise for new treatment of such genetic diseases as cystic fibrosis, muscular dystrophy and hemophilia. But to work well, the new gene-clipping tool must be delivered safely across the cell membrane and into its nucleus, a difficult process that can trigger the cell's defenses and \"trap\" CRISPR\/Cas9, greatly reducing its treatment potential. <\/p>\n
Now, researchers in nanochemistry expert Vincent Rotello's laboratory at the University of Massachusetts Amherst have designed a delivery system using nanoparticles to assist CRISPR\/Cas9 across the membrane and into the cell nucleus while avoiding entrapment by cellular machinery. Details appear in a recent issue of the journal ACS Nano. <\/p>\n
The lab's experiment leader, Rubul Mout, says, \"CRISPR has two components: a scissor-like protein called Cas9, and an RNA molecule called sgRNA that guides Cas9 to its target gene. Once the Cas9-sgRNA pair gets to the destination gene in the nucleus, it can interrogate its genetic mistakes and correct them with the help of the host cell's repair machinery.\" <\/p>\n
He points out that since CRISPR's potential was first discovered in 2012, gene editing or genome engineering has quickly become an intense research topic in biology and medicine. The goal is to treat otherwise incurable genetic diseases by manipulating diseased genes. \"However, to achieve this, biotech and pharmaceutical companies are constantly searching for more efficient CRISPR delivery methods,\" he adds. <\/p>\n
The new delivery method Rotello, Mout and colleagues designed involves engineering the Cas9 protein, named Cas9En, and carrier nanoparticles. Rotello says, \"By finely tuning the interactions between engineered Cas9En protein and nanoparticles, we were able to construct these delivery vectors. The vectors carrying the Cas9 protein and sgRNA come into contact with the cell membrane, fuse, and release the Cas9:sgRNA directly into the cell cytoplasm.\" <\/p>\n
\"Cas9 protein also has a nuclear guiding sequence that ushers the complex into the destination nucleus. The key is to tweak the Cas9 protein,\" he adds. \"We have delivered this Cas9 protein and sgRNA pair into the cell nucleus without getting it trapped on its way. We have watched the delivery process live in real time using sophisticated microscopy.\" <\/p>\n
Mout and colleagues say they can now deliver the Cas9 protein and sgRNA pair into about 90 percent of cells grown in a culture dish with an editing efficiency of about 30 percent. \"Ninety percent cytosolic\/nuclear delivery is a huge improvement compared to others methods,\" Mout points out. <\/p>\n
The researchers believe that the Cas9En may also serve as a platform for delivery of a variety of other materials such as polymers, lipid nanoparticles or self-assembling peptides. Rotello says, \"Now that we have achieved efficient gene editing in cultured cells, we are aiming to edit genes in pre-clinical animal models. We are also interested in gene editing for adoptive therapies, where a diseased cell is isolated from a patient, corrected by CRISPR in the lab, and delivered back to the patient.\" <\/p>\n
Apart from gene editing, the new delivery method may have other uses. For example, another important issue in biology and medicine is tracking DNA and RNA inside cells. Recently, CRISPR has been used to aid in this research. Moumita Ray, another researcher in the Rotello lab, says, \"Our method allows the precise monitoring of Cas9 protein movement inside a cell, opening new opportunities in genomic research.\" <\/p>\n
Explore further: Watching gene editing at work to develop precision therapies <\/p>\n
More information: Rubul Mout et al. Direct Cytosolic Delivery of CRISPR\/Cas9-Ribonucleoprotein for Efficient Gene Editing, ACS Nano (2017). DOI: 10.1021\/acsnano.6b07600<\/p>\n
University of Wisconsin-Madison engineers have developed methods to observe gene editing in action, and they're putting those capabilities to work to improve genetic engineering techniques. <\/p>\n
A study in The Journal of Cell Biology by scientists at the University of Massachusetts Medical School reveals important new details about the inner workings of the CRISPR-Cas9 machinery in live cells that may have implications ... <\/p>\n
The ability to control gene expression in cells allows scientists to understand gene function and manipulate cell fate. Recently, scientists have developed a revolutionary gene-editing tool, called CRIPSR\/Cas9, which employs ... <\/p>\n
Researchers have discovered a way to program cells to inhibit CRISPR-Cas9 activity. \"Anti-CRISPR\" proteins had previously been isolated from viruses that infect bacteria, but now University of Toronto and University of Massachusetts ... <\/p>\n
(Phys.org)A team of researchers with members from several institutions in Japan has developed a new way to edit genes that involves cutting just one strand of DNA rather than both of them, as is normal for CRISPR-Cas9. ... <\/p>\n
Researchers from North Carolina State University and the University of North Carolina at Chapel Hill have for the first time created and used a nanoscale vehicle made of DNA to deliver a CRISPR-Cas9 gene-editing tool into ... <\/p>\n
More and more scientists are using the powerful new gene-editing tool known as CRISPR\/Cas9, a technology isolated from bacteria, that holds promise for new treatment of such genetic diseases as cystic fibrosis, muscular dystrophy ... <\/p>\n
Inspired by the hair of blue tarantulas, researchers from The University of Akron lead a team that made a structural-colored material that shows consistent color from all viewing directions. This finding overturns the conventional ... <\/p>\n
Using tiny snippets of DNA as \"barcodes,\" researchers have developed a new technique for rapidly screening the ability of nanoparticles to selectively deliver therapeutic genes to specific organs of the body. The technique ... <\/p>\n
How the natural defence force within our immune system attacks and destroys harmful invaders such as virus-infected and cancerous cells has been visualised in microscopic detail by scientists from UCL, Birkbeck, University ... <\/p>\n
(Phys.org)In an effort to curb the adverse environmental impacts of paper production, researchers in a new study have developed a light-printable paperpaper that can be printed with UV light, erased by heating to 120 ... <\/p>\n
Scientists used one of the world's most powerful electron microscopes to map the precise location and chemical type of 23,000 atoms in an extremely small particle made of iron and platinum. <\/p>\n
Please sign in to add a comment. Registration is free, and takes less than a minute. Read more <\/p>\n
<\/p>\n
See the article here: February 7, 2017 A cartoon shows gene editing through engineered CRISPR\/Cas9En delivery in the Rotello lab at UMass Amherst. The researchers have overcome an obstacle in the technology by designing a delivery system using nanoparticles to assist CRISPR\/Cas9 across the cell membrane and into the nucleus while avoiding entrapment by cellular machinery. Continue reading
\nOvercoming hurdles in CRISPR gene editing to improve treatment - Phys.Org<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"