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

同源修复模板 DNA 链间交联增强人类细胞基因编辑

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作者:Hannah I Ghasemi, Julien Bacal, Amanda C Yoon, Katherine U Tavasoli, Carmen Cruz, Jonathan T Vu, Brooke M Gardner, Chris D Richardson

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.

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