Show simple item record

dc.contributor.authorAllen, Felicity
dc.contributor.authorCrepaldi, Luca
dc.contributor.authorAlsinet, Clara
dc.contributor.authorStrong, Alexander J
dc.contributor.authorKleshchevnikov, Vitalii
dc.contributor.authorDe Angeli, Pietro
dc.contributor.authorPáleníková, Petra
dc.contributor.authorKhodak, Anton
dc.contributor.authorKiselev, Vladimir
dc.contributor.authorKosicki, Michael
dc.contributor.authorBassett, Andrew R
dc.contributor.authorHarding, Heather
dc.contributor.authorGalanty, Yaron
dc.contributor.authorMuñoz-Martínez, Francisco
dc.contributor.authorMetzakopian, Emmanouil
dc.contributor.authorJackson, Stephen
dc.contributor.authorParts, Leopold
dc.date.accessioned2018-12-14T00:32:07Z
dc.date.available2018-12-14T00:32:07Z
dc.date.issued2018-11-27
dc.identifier.issn1087-0156
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/286946
dc.description.abstractThe DNA mutation produced by cellular repair of a CRISPR-Cas9-generated double-strand break determines its phenotypic effect. It is known that the mutational outcomes are not random, but depend on DNA sequence at the targeted location. Here we systematically study the influence of flanking DNA sequence on repair outcome by measuring the edits generated by >40,000 guide RNAs (gRNAs) in synthetic constructs. We performed the experiments in a range of genetic backgrounds and using alternative CRISPR-Cas9 reagents. In total, we gathered data for >109 mutational outcomes. The majority of reproducible mutations are insertions of a single base, short deletions or longer microhomology-mediated deletions. Each gRNA has an individual cell-line-dependent bias toward particular outcomes. We uncover sequence determinants of the mutations produced and use these to derive a predictor of Cas9 editing outcomes. Improved understanding of sequence repair will allow better design of gene editing experiments.
dc.format.mediumPrint-Electronic
dc.languageeng
dc.publisherSpringer Science and Business Media LLC
dc.titlePredicting the mutations generated by repair of Cas9-induced double-strand breaks.
dc.typeArticle
prism.publicationDate2018
prism.publicationNameNat Biotechnol
dc.identifier.doi10.17863/CAM.34255
dcterms.dateAccepted2018-11-12
rioxxterms.versionofrecord10.1038/nbt.4317
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2018-11-27
dc.contributor.orcidKleshchevnikov, Vitalii [0000-0001-9110-7441]
dc.contributor.orcidKiselev, Vladimir [0000-0003-1893-2255]
dc.contributor.orcidBassett, Andrew R [0000-0003-1632-9137]
dc.contributor.orcidHarding, Heather [0000-0002-7359-7974]
dc.contributor.orcidGalanty, Yaron [0000-0001-7167-9004]
dc.contributor.orcidJackson, Stephen [0000-0001-9317-7937]
dc.identifier.eissn1546-1696
rioxxterms.typeJournal Article/Review
pubs.funder-project-idCancer Research UK (18796)
pubs.funder-project-idWellcome Trust (206388/Z/17/Z)
pubs.funder-project-idWellcome Trust (084812/Z/08/Z)
pubs.funder-project-idWellcome Trust (200848/Z/16/Z)
pubs.funder-project-idWellcome Trust (100140/Z/12/Z)
cam.issuedOnline2018-11-27
rioxxterms.freetoread.startdate2019-05-27


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record