Interstrand crosslinking of homologous repair template DNA enhances gene editing in human cells.

Ghasemi, Hannah I; Bacal, Julien; Yoon, Amanda C; Tavasoli, Katherine U; Cruz, Carmen; Vu, Jonathan T; Gardner, Brooke M; Richardson, Chris D · Nat Biotechnol · 2023

basic_science · Level V

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Abstract

We describe a strategy to boost the efficiency of gene editing via homology-directed repair (HDR) by covalently modifying the template DNA with interstrand crosslinks. Crosslinked templates (xHDRTs) increase Cas9-mediated editing efficiencies by up to fivefold in K562, HEK293T, U2OS, iPS and primary T cells. Increased editing from xHDRTs is driven by events on the template molecule and requires ataxia telangiectasia and Rad3-related (ATR) kinase and components of the Fanconi anemia pathway.

Medical subject headings