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Abudayyeh, Omar O; Gootenberg, Jonathan S; Franklin, Brian; Koob, Jeremy; Kellner, Max J; Ladha, Alim; Joung, Julia; Kirchgatterer, Paul; Cox, David B T; Zhang, Feng
Science (American Association for the Advancement of Science), 07/2019, Volume: 365, Issue: 6451Journal Article
Programmable RNA editing enables reversible recoding of RNA information for research and disease treatment. Previously, we developed a programmable adenosine-to-inosine (A-to-I) RNA editing approach by fusing catalytically inactivate RNA-targeting CRISPR-Cas13 (dCas13) with the adenine deaminase domain of ADAR2. Here, we report a cytidine-to-uridine (C-to-U) RNA editor, referred to as RNA Editing for Specific C-to-U Exchange (RESCUE), by directly evolving ADAR2 into a cytidine deaminase. RESCUE doubles the number of mutations targetable by RNA editing and enables modulation of phosphosignaling-relevant residues. We apply RESCUE to drive β-catenin activation and cellular growth. Furthermore, RESCUE retains A-to-I editing activity, enabling multiplexed C-to-U and A-to-I editing through the use of tailored guide RNAs.
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