Fiche publication
Date publication
juillet 2025
Journal
Nature biotechnology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr CULLOT Grégoire
Tous les auteurs :
Karasu ME, Toufektchan E, Chen Y, Albertelli A, Cullot G, Maciejowski J, Corn JE
Lien Pubmed
Résumé
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.
Mots clés
Humans, Exodeoxyribonucleases, genetics, CRISPR-Cas Systems, genetics, Phosphoproteins, genetics, Homologous Recombination, genetics, Gene Editing, methods
Référence
Nat Biotechnol. 2025 07;43(7):1168-1176
