Scientists on the Florida campus of The Scripps Research Institute (TSRI) have improved a state-of-the-art gene-editing technology to advance the system’s ability to target, cut and paste genes within human and animal cells—and broadening the ways the CRISPR-Cpf1 editing system may be used to study and fight human diseases.
Professor Michael Farzan, co-chair of TSRI’s Department of Immunology and Microbiology, and TSRI Research Associate Guocai Zhong improved the efficiency of the CRISPR-Cpf1 gene editing system by incorporating guide RNAs with “multiplexing” capability. Guide RNAs are short nucleic acid strings that lead the CRISPR molecular scissors to their intended gene targets. The TSRI discovery means multiple genetic targets in a cell may be hit by each CRISPR-Cpf1 complex.
“This system simplifies and significantly improves the efficiency of simultaneous editing of multiple genes, or multiple sites of a single gene,” Zhong said. “This could be very useful when multiple disease-related genes or multiple sites of a disease-related gene need to be targeted.”
“This approach improves gene editing for a number of applications,” Farzan added. “The system makes some applications more efficient and other applications possible.”
This study was published as an advanced online paper in the journal Nature Chemical Biology on June 19, 2017.
TSRI Advance Makes CRISPR More Efficient
Short for “Clustered Regularly Interspaced Short Palindromic Repeat,” the CRISPR gene editing system exploits an ancient bacterial immune defense process. Some microbes thwart viral infection by sequestering a piece of a virus’ foreign genetic material within its own DNA, to serve as a template. The next time the viral sequence is encountered by the microbe, it’s recognized immediately and cut up for disposal with the help of two types of RNA. Molecules called guide RNAs provide the map to the invader, and CRISPR effector proteins act as the scissors that cut it apart.
Over the last five years, the CRISPR gene editing system has revolutionized microbiology and renewed hopes that genetic engineering might eventually become a useful treatment for disease. But time has revealed the technology’s limitations. For one, gene therapy currently requires using a viral shell to serve as the delivery package for the therapeutic genetic material. The CRISPR molecule is simply too large to fit with multiple guide RNAs into the most popular and useful viral packaging system.
The new study from Farzan and colleagues helps solve this problem by letting scientists package multiple guide RNAs.
This advance could be important if gene therapy is to treat diseases such as hepatitis B, Farzan said. After infection, hepatitis B DNA sits in liver cells, slowly directing the production of new viruses, ultimately leading to liver damage, cirrhosis and even cancer. The improved CRISPR-Cpf1 system, with its ability to ‘multiplex,’ could more efficiently digest the viral DNA, before the liver is irrevocably damaged, he said.
“Efficiency is important. If you modify 25 cells in the liver, it is meaningless. But if you modify half the cells in the liver, that is powerful,” Farzan said. “There are other good cases—say muscular dystrophy—where if you can repair the gene in enough muscle cells, you can restore the muscle function.”
Two types of these molecular scissors are now being widely used for gene editing purposes: Cas9 and Cpf1. Farzan said he focused on Cpf1 because it is more precise in mammalian cells. The Cpf1 molecule they studied was sourced from two types of bacteria, Lachnospiraceae bacterium and Acidaminococus sp., whose activity has been previously studied in E. coli. A key property of these molecules is they are able to grab their guide RNAs out of a long string of such RNA; but it was not clear that it would work with RNA produced from mammalian cells. Guocai tested this idea by editing a firefly bioluminescence gene into the cell’s chromosome. The modified CRISPR-Cpf1 system worked as anticipated.
“This means we can use simpler delivery systems for directing the CRISPR effector protein plus guide RNAs,” Farzan said. “It’s going to make the CRISPR process more efficient for a variety of applications.”
Looking forward, Farzan said the Cpf1 protein needs to be more broadly understood so that its utility in delivering gene therapy vectors can be further expanded.
The Latest on: CRISPR-Cpf1
- Better Buy: Editas Medicine, Inc. vs. CRISPR Therapeutics on September 7, 2018 at 5:18 am
Second, I like that Editas has no competition in LCA10 or the other genetic eye diseases that it's targeting. Third, the biotech's CRISPR-Cpf1 program could give it an advantage in addressing some gen... […]
- Editas Medicine Announces Transition of Chief Medical Officer on August 26, 2018 at 5:00 pm
About Editas Medicine As a leading genome editing company, Editas Medicine is focused on translating the power and potential of the CRISPR/Cas9 and CRISPR/Cpf1 genome editing systems into a robust pip... […]
- Researchers use CRISPR to edit DNA outside of the cell for the first time on April 19, 2018 at 7:00 am
Bringing the CRISPR Revolution to Everyday Cancer Care The Gene Editing Institute is now working to use the new CRISPR-Cpf1 breakthrough to provide something akin to CRISPR on a computer chip to a com... […]
- Editas Medicine Reports Data Supporting Multiple Approaches to Create Treatments for Sickle Cell Disease and Beta-Thalassemia on December 11, 2017 at 3:02 pm
In these proof of concept experiments, the Company demonstrated CRISPR/Cas9 homology directed repair (HDR) and CRISPR/Cpf1-directed editing in human CD34 + cells. The Company reported these data today ... […]
- Future CRISPR Proteins Provide A Moat To Editas Medicine on October 27, 2017 at 6:40 am
EDIT may see itself able to develop a moat over the next 5 years as they continue to explore the newly discovered CRISPR-Cpf1 by Zhang, AT to GC Cas9 developed by Liu and a Cas13 analog for editing RN... […]
- Firefly gene illuminates ability of optimized CRISPR-Cpf1 to efficiently edit human genome on June 19, 2017 at 8:02 am
Professor Michael Farzan, co-chair of TSRI's Department of Immunology and Microbiology. Credit: The Scripps Research Institute Scientists on the Florida campus of The Scripps Research Institute (TSRI) ... […]
- Firefly Gene Illuminates Ability of Optimized CRISPR-Cpf1 to Efficiently Edit Human Genome on June 17, 2017 at 9:00 am
Newswise — JUPITER, FL – June 19, 2017 – Scientists on the Florida campus of The Scripps Research Institute (TSRI) have improved a state-of-the-art gene-editing technology to advance the system’s abil... […]
- A New Flavor of CRISPR Could Tackle Some of the Worst Genetic Diseases on April 13, 2017 at 3:04 am
Cardiomyocytes from patients with Duchenne muscular dystrophy corrected by CRISPR-Cpf1. Image: Science Advances Most of the time, when people talk about the cutting edge gene editing technology CRISPR ... […]
- Gene-editing alternative corrects Duchenne muscular dystrophy on April 11, 2017 at 5:00 pm
Using the new gene-editing enzyme CRISPR-Cpf1, researchers at UT Southwestern Medical Center have successfully corrected Duchenne muscular dystrophy in human cells and mice in the lab. The UT Southwes... […]
- Editas inks deal with Allergan for as many as five gene-editing ocular programs; shares ahead 10% premarket on March 14, 2017 at 6:13 am
The agreement includes programs based on the CRISPR/Cas9 platform as well as CRISPR/Cpf1. Under the terms of the deal, Editas will receive an upfront payment of $90M, milestones and sales-based royalt... […]
via Google News and Bing News