CLUSTER guide RNAs enable precise and efficient RNA editing with endogenous ADAR enzymes in vivo.
Journal
Nature biotechnology
ISSN: 1546-1696
Titre abrégé: Nat Biotechnol
Pays: United States
ID NLM: 9604648
Informations de publication
Date de publication:
05 2022
05 2022
Historique:
received:
27
10
2020
accepted:
23
09
2021
pubmed:
5
1
2022
medline:
20
5
2022
entrez:
4
1
2022
Statut:
ppublish
Résumé
RNA base editing represents a promising alternative to genome editing. Recent approaches harness the endogenous RNA-editing enzyme adenosine deaminase acting on RNA (ADAR) to circumvent problems caused by ectopic expression of engineered editing enzymes, but suffer from sequence restriction, lack of efficiency and bystander editing. Here we present in silico-optimized CLUSTER guide RNAs that bind their target messenger RNAs in a multivalent fashion, achieve editing with high precision and efficiency and enable targeting of sequences that were not accessible using previous gRNA designs. CLUSTER gRNAs can be genetically encoded and delivered using viruses, and are active in a wide range of cell lines. In cell culture, CLUSTER gRNAs achieve on-target editing of endogenous transcripts with yields of up to 45% without bystander editing. In vivo, CLUSTER gRNAs delivered to mouse liver by hydrodynamic tail vein injection edited reporter constructs at rates of up to 10%. The CLUSTER approach opens avenues for drug development in the field of RNA base editing.
Identifiants
pubmed: 34980913
doi: 10.1038/s41587-021-01105-0
pii: 10.1038/s41587-021-01105-0
doi:
Substances chimiques
RNA, Guide
0
RNA, Messenger
0
RNA
63231-63-0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
759-768Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.
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