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dc.contributor.authorBowden, Anne
dc.contributor.authorJuarez, David A Morales
dc.contributor.authorSczaniecka-Clift, Matylda
dc.contributor.authorAgudo, Maria Martin
dc.contributor.authorLukashchuk, Natalia
dc.contributor.authorThomas, John
dc.contributor.authorJackson, Stephen
dc.date.accessioned2020-04-24T23:30:44Z
dc.date.available2020-04-24T23:30:44Z
dc.date.issued2020-02-20
dc.identifier.issn2050-084X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/304696
dc.description.abstract<jats:title>Abstract</jats:title><jats:p>CRISPR-Cas9 genome engineering has revolutionised high-throughput functional genomic screens. However, recent work has raised concerns regarding the performance of CRISPR-Cas9 screens using <jats:italic>TP53</jats:italic> wild-type human cells due to a p53-mediated DNA damage response (DDR) limiting the efficiency of generating viable edited cells. To directly assess the impact of cellular p53 status on CRISPR-Cas9 screen performance, we carried out parallel CRISPR-Cas9 screens in wild-type and <jats:italic>TP53</jats:italic> knockout human retinal pigment epithelial cells using a focused dual guide RNA library targeting 852 DDR-associated genes. Our work demonstrates that although functional p53 status negatively affects identification of significantly depleted genes, optimal screen design can nevertheless enable robust screen performance. Through analysis of our own and published screen data, we highlight key factors for successful screens in both wild-type and p53-deficient cells.</jats:p>
dc.publishereLife Sciences Publications Ltd
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleParallel CRISPR-Cas9 screens clarify impacts of p53 on screen performance
dc.typeArticle
prism.publicationNameeLife
dc.identifier.doi10.17863/CAM.51776
dcterms.dateAccepted2020-04-23
rioxxterms.versionofrecord10.7554/eLife.55325
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2020-04-23
dc.contributor.orcidBowden, Anne [0000-0003-1138-4452]
dc.contributor.orcidThomas, John [0000-0003-2425-8412]
dc.contributor.orcidJackson, Stephen [0000-0001-9317-7937]
dc.identifier.eissn2050-084X
rioxxterms.typeJournal Article/Review
cam.issuedOnline2020-05-22
cam.orpheus.successMon Jul 20 07:54:57 BST 2020 - The item has an open VoR version.
cam.orpheus.counter11
rioxxterms.freetoread.startdate2100-01-01


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International