RNA editing with CRISPR-Cas13.

Science
Authors
Keywords
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.

Year of Publication
2017
Journal
Science
Volume
358
Issue
6366
Pages
1019-1027
Date Published
2017 11 24
ISSN
1095-9203
DOI
10.1126/science.aaq0180
PubMed ID
29070703
PubMed Central ID
PMC5793859
Links
Grant list
R01 MH110049 / MH / NIMH NIH HHS / United States
T32 GM007753 / GM / NIGMS NIH HHS / United States
DP1 HL141201 / HL / NHLBI NIH HHS / United States
R01 HG009761 / HG / NHGRI NIH HHS / United States
RM1 HG006193 / HG / NHGRI NIH HHS / United States
F30 CA210382 / CA / NCI NIH HHS / United States