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

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

doi:10.1038/s41587-022-01654-y

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

Nat Biotechnol. 2023 Oct;41(10):1398-1404. doi: 10.1038/s41587-022-01654-y. Epub 2023 Feb 27.

Authors

Hannah I Ghasemi¹, Julien Bacal¹, Amanda C Yoon¹, Katherine U Tavasoli¹, Carmen Cruz¹, Jonathan T Vu¹, Brooke M Gardner¹, Chris D Richardson²

Affiliations

¹ Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA, USA.
² Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA, USA. chrisrichardson@ucsb.edu.

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.

MeSH terms

CRISPR-Cas Systems / genetics
Cross-Linking Reagents / chemistry
DNA Repair / genetics
DNA* / chemistry
DNA* / genetics
Gene Editing* / methods
HEK293 Cells
Humans
Recombinational DNA Repair
Templates, Genetic

Substances

DNA
Cross-Linking Reagents

Abstract

MeSH terms

Substances

Grants and funding