Abstract
Prime editors have a broad range of potential research and clinical applications. However, methods to delineate their genome-wide editing activities have generally relied on indirect genome-wide editing assessments or the computational prediction of near-cognate sequences. Here we describe a genome-wide approach for the identification of potential prime editor off-target sites, which we call PE-tag. This method relies on the attachment or insertion of an amplification tag at sites of prime editor activity to allow their identification. PE-tag enables genome-wide profiling of off-target sites in vitro using extracted genomic DNA, in mammalian cell lines and in the adult mouse liver. PE-tag components can be delivered in a variety of formats for off-target site detection. Our studies are consistent with the high specificity previously described for prime editor systems, but we find that off-target editing rates are influenced by prime editing guide RNA design. PE-tag represents an accessible, rapid and sensitive approach for the genome-wide identification of prime editor activity and the evaluation of prime editor safety.
Original language | English (US) |
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Pages (from-to) | 898-907 |
Number of pages | 10 |
Journal | Nature Methods |
Volume | 20 |
Issue number | 6 |
DOIs | |
State | Published - Jun 2023 |
Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2023, The Author(s), under exclusive licence to Springer Nature America, Inc.
PubMed: MeSH publication types
- Journal Article