RNA editing with CRISPR-Cas13
Name
nihms920681.pdf
Description
Accepted version
Size
2.48 MB
Format
Adobe PDF
Checksum (MD5)
44cea0cccfb8407f36e7593ce4b455cb
Author(s)
Cox, David Benjamin Turitz
Gootenberg, Jonathan S.
Abudayyeh, Omar O.
Franklin, Brian
Kellner, Max J.
Joung, Julia
Zhang, Feng
Date Issued
October 2017
Publisher
American Association for the Advancement of Science (AAAS)
Citation
Cox, David B. T. et al. "RNA editing with CRISPR-Cas13." Science 358, 6366 (24 Nov 2017): 1019-1027 ©2017, American Association for the Advancement of Science.
Version
Author's final manuscript
Abstract
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.
MIT Department
Broad Institute of MIT and Harvard
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Harvard University--MIT Division of Health Sciences and Technology
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1126/science.aaq0180
