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Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells

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dc.contributor.authorKim, Daesik-
dc.contributor.authorBae, Sangsu-
dc.contributor.authorPark, Jeongbin-
dc.contributor.authorKim, Eunji-
dc.contributor.authorKim, Seokjoong-
dc.contributor.authorYu, Hye Ryeong-
dc.contributor.authorHwang, Jinha-
dc.contributor.authorKim, Jong-Il-
dc.contributor.authorKim, Jin-Soo-
dc.date.accessioned2020-04-27T12:43:30Z-
dc.date.available2020-04-27T12:43:30Z-
dc.date.issued2015-03-
dc.identifier.citationNature Methods, Vol.12 No.3, pp.237-243-
dc.identifier.issn1548-7091-
dc.identifier.other57993-
dc.identifier.urihttps://hdl.handle.net/10371/165651-
dc.description.abstractAlthough RNA-guided genome editing via the CRISPR-Cas9 system is now widely used in biomedical research, genome-wide target specificities of Cas9 nucleases remain controversial. Here we present Digenome-seq, in vitro Cas9-digested whole-genome sequencing, to profile genome-wide Cas9 off-target effects in human cells. This in vitro digest yields sequence reads with the same 5' ends at cleavage sites that can be computationally identified. We validated off-target sites at which insertions or deletions were induced with frequencies below 0.1%, near the detection limit of targeted deep sequencing. We also showed that Cas9 nucleases can be highly specific, inducing off-target mutations at merely several, rather than thousands of, sites in the entire genome and that Cas9 off-target effects can be avoided by replacing 'promiscuous' single guide RNAs (sgRNAs) with modified sgRNAs. Digenome-seq is a robust, sensitive, unbiased and cost-effective method for profiling genome-wide off-target effects of programmable nucleases including Cas9.-
dc.titleDigenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells-
dc.typeArticle-
dc.contributor.AlternativeAuthor김종일-
dc.contributor.AlternativeAuthor김진수-
dc.identifier.doi10.1038/NMETH.3284-
dc.citation.journaltitleNature Methods-
dc.identifier.scopusid2-s2.0-84923846574-
dc.citation.endpage243-
dc.citation.number3-
dc.citation.startpage237-
dc.citation.volume12-
dc.identifier.urlhttps://www.nature.com/articles/nmeth.3284-
dc.identifier.rimsid57993-
dc.identifier.sci000350670300025-
dc.description.isOpenAccessN-
dc.contributor.affiliatedAuthorKim, Jin-Soo-
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