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Precise in vivo RNA base editing with a wobble-enhanced circular CLUSTER guide RNA.
- Source :
-
Nature biotechnology [Nat Biotechnol] 2024 Jul 12. Date of Electronic Publication: 2024 Jul 12. - Publication Year :
- 2024
- Publisher :
- Ahead of Print
-
Abstract
- Recruiting the endogenous editing enzyme adenosine deaminase acting on RNA (ADAR) with tailored guide RNAs for adenosine-to-inosine (A-to-I) RNA base editing is promising for safely manipulating genetic information at the RNA level. However, the precision and efficiency of editing are often compromised by bystander off-target editing. Here, we find that in 5'-UAN triplets, which dominate bystander editing, G•U wobble base pairs effectively mitigate off-target events while maintaining high on-target efficiency. This strategy is universally applicable to existing A-to-I RNA base-editing systems and complements other suppression methods such as G•A mismatches and uridine (U) depletion. Combining wobble base pairing with a circularized format of the CLUSTER approach achieves highly precise and efficient editing (up to 87%) of a disease-relevant mutation in the Mecp2 transcript in cell culture. Virus-mediated delivery of the guide RNA alone realizes functional MeCP2 protein restoration in the central nervous system of a murine Rett syndrome model with editing yields of up to 19% and excellent bystander control in vivo.<br /> (© 2024. The Author(s).)
Details
- Language :
- English
- ISSN :
- 1546-1696
- Database :
- MEDLINE
- Journal :
- Nature biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 38997581
- Full Text :
- https://doi.org/10.1038/s41587-024-02313-0