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Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Daesik | - |
dc.contributor.author | Bae, Sangsu | - |
dc.contributor.author | Park, Jeongbin | - |
dc.contributor.author | Kim, Eunji | - |
dc.contributor.author | Kim, Seokjoong | - |
dc.contributor.author | Yu, Hye Ryeong | - |
dc.contributor.author | Hwang, Jinha | - |
dc.contributor.author | Kim, Jong-Il | - |
dc.contributor.author | Kim, Jin-Soo | - |
dc.date.accessioned | 2020-04-27T12:43:30Z | - |
dc.date.available | 2020-04-27T12:43:30Z | - |
dc.date.created | 2018-10-04 | - |
dc.date.created | 2018-10-04 | - |
dc.date.created | 2018-10-04 | - |
dc.date.issued | 2015-03 | - |
dc.identifier.citation | Nature Methods, Vol.12 No.3, pp.237-243 | - |
dc.identifier.issn | 1548-7091 | - |
dc.identifier.other | 57993 | - |
dc.identifier.uri | https://hdl.handle.net/10371/165651 | - |
dc.description.abstract | Although 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.language | 영어 | - |
dc.publisher | Nature Publishing Group | - |
dc.title | Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells | - |
dc.type | Article | - |
dc.contributor.AlternativeAuthor | 김종일 | - |
dc.contributor.AlternativeAuthor | 김진수 | - |
dc.identifier.doi | 10.1038/NMETH.3284 | - |
dc.citation.journaltitle | Nature Methods | - |
dc.identifier.wosid | 000350670300025 | - |
dc.identifier.scopusid | 2-s2.0-84923846574 | - |
dc.citation.endpage | 243 | - |
dc.citation.number | 3 | - |
dc.citation.startpage | 237 | - |
dc.citation.volume | 12 | - |
dc.identifier.sci | 000350670300025 | - |
dc.description.isOpenAccess | N | - |
dc.contributor.affiliatedAuthor | Kim, Jong-Il | - |
dc.contributor.affiliatedAuthor | Kim, Jin-Soo | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.subject.keywordPlus | ZINC-FINGER NUCLEASES | - |
dc.subject.keywordPlus | CRISPR/CAS9 SYSTEMS | - |
dc.subject.keywordPlus | GUIDE RNA | - |
dc.subject.keywordPlus | CAS9 | - |
dc.subject.keywordPlus | SPECIFICITY | - |
dc.subject.keywordPlus | ENDONUCLEASE | - |
dc.subject.keywordPlus | DNA | - |
dc.subject.keywordPlus | RIBONUCLEOPROTEINS | - |
dc.subject.keywordPlus | MUTATIONS | - |
dc.subject.keywordPlus | NICKASES | - |
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