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Nat Commun ; 13(1): 760, 2022 02 09.
Article in English | MEDLINE | ID: mdl-35140211

ABSTRACT

Prime editing (PE) is a powerful genome engineering approach that enables the introduction of base substitutions, insertions and deletions into any given genomic locus. However, the efficiency of PE varies widely and depends not only on the genomic region targeted, but also on the genetic background of the edited cell. Here, to determine which cellular factors affect PE efficiency, we carry out a focused genetic screen targeting 32 DNA repair factors, spanning all reported repair pathways. We show that, depending on cell line and type of edit, ablation of mismatch repair (MMR) affords a 2-17 fold increase in PE efficiency, across several human cell lines, types of edits and genomic loci. The accumulation of the key MMR factors MLH1 and MSH2 at PE sites argues for direct involvement of MMR in PE control. Our results shed new light on the mechanism of PE and suggest how its efficiency might be optimised.


Subject(s)
DNA Mismatch Repair , Gene Editing , Cell Line , DNA Repair , DNA-Binding Proteins , Genetic Testing , HEK293 Cells , Humans , MutL Protein Homolog 1/genetics , MutS Homolog 2 Protein/metabolism , Mutation, Missense
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