RNA editing with CRISPR-Cas13

• Main Authors: Cox, David Benjamin Turitz (Author), Gootenberg, Jonathan S. (Author), Abudayyeh, Omar O. (Author), Franklin, Brian (Author), Kellner, Max J. (Author), Joung, Julia (Author), Zhang, Feng (Author)
• Other Authors: Broad Institute of MIT and Harvard (Contributor), McGovern Institute for Brain Research at MIT (Contributor), Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences (Contributor), Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Biology (Contributor), Harvard University- (Contributor)
• Format: Article
• Language: English
• Published: American Association for the Advancement of Science (AAAS), 2020-05-07T12:51:49Z.
• Subjects: Article
• Online Access: Get fulltext

Nucleic acid editing holds promise for treating genetic disease, particularly at the RNA level, where disease-relevant sequences can be rescued to yield functional protein products. Type VI CRISPR-Cas systems contain the programmable single-effector RNA-guided ribonuclease Cas13. We profiled type VI systems in order to engineer a Cas13 ortholog capable of robust knockdown and demonstrated RNA editing by using catalytically inactive Cas13 (dCas13) to direct adenosine-to-inosine deaminase activity by ADAR2 (adenosine deaminase acting on RNA type 2) to transcripts in mammalian cells. This system, referred to as RNA Editing for Programmable A to I Replacement (REPAIR), which has no strict sequence constraints, can be used to edit full-length transcripts containing pathogenic mutations. We further engineered this system to create a high-specificity variant and minimized the system to facilitate viral delivery. REPAIR presents a promising RNA-editing platform with broad applicability for research, therapeutics, and biotechnology.
National Institute of Allergy and Infectious Diseases (U.S.) (Grant R01AI117043)
United States. Air Force. Office of Scientific Research (Grant FA9550-14-1-0060)
National Institute of Mental Health (U.S.) (Grant 5DP1-MH100706)
National Institute of Mental Health (U.S.) (Grant 1R01-MH110049)