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A cytosine deaminase for programmable single-base RNA editing.

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 ?-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.

SUBMITTER: Abudayyeh OO

PROVIDER: S-EPMC6956565 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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A cytosine deaminase for programmable single-base RNA editing.

Abudayyeh Omar O OO Gootenberg Jonathan S JS Franklin Brian B Koob Jeremy J Kellner Max J MJ Ladha Alim A Joung Julia J Kirchgatterer Paul P Cox David B T DBT Zhang Feng F

Science (New York, N.Y.) 20190711 6451

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 ...[more]

PMID: 31296651

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