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Science. 2019 Jul 26;365(6451):382-386. doi: 10.1126/science.aax7063. Epub 2019 Jul 11.

A cytosine deaminase for programmable single-base RNA editing.

Author information

1
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
2
McGovern Institute for Brain Research at MIT, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
3
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
4
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. zhang@broadinstitute.org.
5
Department of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
#
Contributed equally

Abstract

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.

PMID:
31296651
DOI:
10.1126/science.aax7063

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