Removal of TREX1 activity enhances CRISPR-Cas9-mediated homologous recombination Journal Article

   


Authors: Karasu, M. E.; Toufektchan, E.; Chen, Y. Y.; Albertelli, A.; Cullot, G.; Maciejowski, J.; Corn, J. E.
Article Title: Removal of TREX1 activity enhances CRISPR-Cas9-mediated homologous recombination
Abstract: CRISPR-Cas9-mediated homology-directed repair (HDR) can introduce desired mutations at targeted genomic sites, but achieving high efficiencies is a major hurdle in many cell types, including cells deficient in DNA repair activity. In this study, we used genome-wide screening in Fanconi anemia patient lymphoblastic cell lines to uncover suppressors of CRISPR-Cas9-mediated HDR. We found that a single exonuclease, TREX1, reduces HDR efficiency when the repair template is a single-stranded or linearized double-stranded DNA. TREX1 expression serves as a biomarker for CRISPR-Cas9-mediated HDR in that the high TREX1 expression present in many different cell types (such as U2OS, Jurkat, MDA-MB-231 and primary T cells as well as hematopoietic stem and progenitor cells) predicts poor HDR. Here we demonstrate rescue of HDR efficiency (ranging from two-fold to eight-fold improvement) either by TREX1 knockout or by the use of single-stranded DNA templates chemically protected from TREX1 activity. Our data explain why some cell types are easier to edit than others and indicate routes for increasing CRISPR-Cas9-mediated HDR in TREX1-expressing contexts. Homologous recombination in CRISPR-Cas9 genome editing is increased by blocking an exonuclease activity.
Keywords: gene; mutations; expression; cells; activation; dna-repair; fanconi-anemia; stem; lupus
Journal Title: Nature Biotechnology
Volume: 43
Issue: 7
ISSN: 1087-0156
Publisher: Nature Publishing Group  
Date Published: 2025-07-01
Language: English
ACCESSION: WOS:001289935000002
DOI: 10.1038/s41587-024-02356-3
PROVIDER: wos
PMCID: PMC12263433
PUBMED: 39134754
Notes: Article -- Source: Wos
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