Show simple item record

dc.contributor.authorHartenian, E.
dc.contributor.authorShi, X.
dc.contributor.authorMikkelsen, T. S.
dc.contributor.authorHeckl, D.
dc.contributor.authorEbert, B. L.
dc.contributor.authorRoot, D. E.
dc.contributor.authorDoench, J. G.
dc.contributor.authorShalem, Ophir
dc.contributor.authorSanjana, Neville E
dc.contributor.authorScott, David Arthur
dc.contributor.authorZhang, Feng
dc.date.accessioned2017-09-15T20:18:35Z
dc.date.available2017-09-15T20:18:35Z
dc.date.issued2014-01
dc.date.submitted2013-10
dc.identifier.issn0036-8075
dc.identifier.issn1095-9203
dc.identifier.urihttp://hdl.handle.net/1721.1/111576
dc.description.abstractThe simplicity of programming the CRISPR (clustered regularly interspaced short palindromic repeats)–associated nuclease Cas9 to modify specific genomic loci suggests a new way to interrogate gene function on a genome-wide scale. We show that lentiviral delivery of a genome-scale CRISPR-Cas9 knockout (GeCKO) library targeting 18,080 genes with 64,751 unique guide sequences enables both negative and positive selection screening in human cells. First, we used the GeCKO library to identify genes essential for cell viability in cancer and pluripotent stem cells. Next, in a melanoma model, we screened for genes whose loss is involved in resistance to vemurafenib, a therapeutic RAF inhibitor. Our highest-ranking candidates include previously validated genes NF1 and MED12, as well as novel hits NF2, CUL3, TADA2B, and TADA1. We observe a high level of consistency between independent guide RNAs targeting the same gene and a high rate of hit confirmation, demonstrating the promise of genome-scale screening with Cas9.en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Award 1DP1-MH100706)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (1R01-DK097768)en_US
dc.language.isoen_US
dc.publisherAmerican Association for the Advancement of Science (AAAS)en_US
dc.relation.isversionofhttp://dx.doi.org/10.1126/science.1247005en_US
dc.rightsCreative Commons Attribution-Noncommercial-Share Alikeen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/en_US
dc.sourcePMCen_US
dc.titleGenome-Scale CRISPR-Cas9 Knockout Screening in Human Cellsen_US
dc.typeArticleen_US
dc.identifier.citationShalem, O. "Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells." Science 343, 6166 (January 2014): 84-87 © 2014 American Association for the Advancement of Science (AAAS)en_US
dc.contributor.departmentMcGovern Institute for Brain Research at MITen_US
dc.contributor.mitauthorShalem, Ophir
dc.contributor.mitauthorSanjana, Neville E
dc.contributor.mitauthorScott, David Arthur
dc.contributor.mitauthorZhang, Feng
dc.relation.journalScienceen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsShalem, O.; Sanjana, N. E.; Hartenian, E.; Shi, X.; Scott, D. A.; Mikkelsen, T. S.; Heckl, D.; Ebert, B. L.; Root, D. E.; Doench, J. G.; Zhang, F.en_US
dspace.embargo.termsNen_US
dc.identifier.orcidhttps://orcid.org/0000-0002-2639-9879
dc.identifier.orcidhttps://orcid.org/0000-0003-2782-2509
mit.licenseOPEN_ACCESS_POLICYen_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record