The RESCUE RNA editor enables programmable cytidine-to-uridine (C-to-U) RNA editing, doubling targetable mutations and allowing multiplexed C-to-U and A-to-I editing.
The development of RESCUE expands programmable RNA editing capabilities to include C-to-U exchanges, enabling broader research and potential therapeutic applications.
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.
Abudayyeh et al. (2019) studied this question. RESCUE (RNA Editing for Specific C-to-U Exchange) was evaluated on C-to-U RNA editing and modulation of phosphosignaling-relevant residues. The RESCUE RNA editor enables programmable cytidine-to-uridine (C-to-U) RNA editing, doubling targetable mutations and allowing multiplexed C-to-U and A-to-I editing.