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dc.contributor.authorKosicki, Michael
dc.contributor.authorAllen, Felicity
dc.contributor.authorSteward, Frances
dc.contributor.authorTomberg, Kärt
dc.contributor.authorPan, Yangyang
dc.contributor.authorBradley, Allan
dc.date.accessioned2022-06-14T16:01:05Z
dc.date.available2022-06-14T16:01:05Z
dc.date.issued2022-06-14
dc.date.submitted2020-08-05
dc.identifier.issn2041-1723
dc.identifier.others41467-022-30480-8
dc.identifier.other30480
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/338093
dc.description.abstractRepair of Cas9-induced double-stranded breaks results primarily in formation of small insertions and deletions (indels), but can also cause potentially harmful large deletions. While mechanisms leading to the creation of small indels are relatively well understood, very little is known about the origins of large deletions. Using a library of clonal NGS-validated mouse embryonic stem cells deficient for 32 DNA repair genes, we have shown that large deletion frequency increases in cells impaired for non-homologous end joining and decreases in cells deficient for the central resection gene Nbn and the microhomology-mediated end joining gene Polq. Across deficient clones, increase in large deletion frequency was closely correlated with the increase in the extent of microhomology and the size of small indels, implying a continuity of repair processes across different genomic scales. Furthermore, by targeting diverse genomic sites, we identified examples of repair processes that were highly locus-specific, discovering a role for exonuclease Trex1. Finally, we present evidence that indel sizes increase with the overall efficiency of Cas9 mutagenesis. These findings may have impact on both basic research and clinical use of CRISPR-Cas9, in particular in conjunction with repair pathway modulation.
dc.languageen
dc.publisherSpringer Science and Business Media LLC
dc.subjectArticle
dc.subject/631/337/4041/3196
dc.subject/631/337/1427/2122
dc.subjectarticle
dc.titleCas9-induced large deletions and small indels are controlled in a convergent fashion.
dc.typeArticle
dc.date.updated2022-06-14T16:01:04Z
prism.issueIdentifier1
prism.publicationNameNat Commun
prism.volume13
dc.identifier.doi10.17863/CAM.85502
dcterms.dateAccepted2022-04-28
rioxxterms.versionofrecord10.1038/s41467-022-30480-8
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
dc.contributor.orcidTomberg, Kärt [0000-0002-6144-3601]
dc.contributor.orcidBradley, Allan [0000-0002-2349-8839]
dc.identifier.eissn2041-1723
pubs.funder-project-idWellcome Trust (Wellcome) (098051)
cam.issuedOnline2022-06-14


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