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

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.

SUBMITTER: Ghasemi HI

PROVIDER: S-EPMC10460463 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature

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

Ghasemi Hannah I HI Bacal Julien J Yoon Amanda C AC Tavasoli Katherine U KU Cruz Carmen C Vu Jonathan T JT Gardner Brooke M BM Richardson Chris D CD

Nature biotechnology 20230227 10

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. ...[more]

PMID: 36849829

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BRCA1 functions independently of homologous recombination in DNA interstrand crosslink repair

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Complementation of hypersensitivity to DNA interstrand crosslinking agents demonstrates that XRCC2 is a Fanconi anaemia gene.

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| S-EPMC5035190 | biostudies-literature

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