RNA editing with CRISPR-Cas13

DBT Cox, JS Gootenberg, OO Abudayyeh, B Franklin… - Science, 2017 - science.org
Science, 2017science.org
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 …
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.
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