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유전체교정연구단
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Genome-wide target specificity of CRISPR RNA-guided adenine base editors

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dc.contributor.authorDae Sik Kim-
dc.contributor.authorDa-eun Kim-
dc.contributor.authorGyeorae Lee-
dc.contributor.authorSung-Ik Cho-
dc.contributor.authorJin Soo Kim-
dc.date.available2019-08-19T02:06:28Z-
dc.date.created2019-03-19-
dc.date.issued2019-04-
dc.identifier.issn1087-0156-
dc.identifier.urihttps://pr.ibs.re.kr/handle/8788114/5993-
dc.description.abstractAdenine base editors 1 enable efficient targeted adenine-to-guanine single nucleotide conversions to induce or correct point mutations in human cells, animals, and plants 1–4 . Here we present a modified version of Digenome-seq, an in vitro method for identifying CRISPR (clustered regularly interspaced short palindromic repeats)-induced double-strand breaks using whole-genome sequencing 5–8 , to assess genome-wide target specificity of adenine base editors. To produce double-strand breaks at sites containing inosines, the products of adenine deamination, we treat human genomic DNA with an adenine base editor 7.10 protein–guide RNA complex and either endonuclease V or a combination of human alkyladenine DNA glycosylase and endonuclease VIII in vitro. Digenome-seq detects adenine base editor off-target sites with a substitution frequency of 0.1% or more. We show that adenine base editor 7.10, the cytosine base editor BE3, and unmodified CRISPR-associated protein 9 (Cas9) often recognize different off-target sites, highlighting the need for independent assessments of their genome-wide specificities 6 . Using targeted sequencing, we also show that use of preassembled adenine base editor ribonucleoproteins, modified guide RNAs 5,8–11 , and Sniper/Cas9 (ref. 12 ) reduces adenine base editor off-target activity in human cells. © 2019, The Author(s), under exclusive licence to Springer Nature America, Inc-
dc.description.uri1-
dc.language영어-
dc.publisherNATURE PUBLISHING GROUP-
dc.titleGenome-wide target specificity of CRISPR RNA-guided adenine base editors-
dc.typeArticle-
dc.type.rimsART-
dc.identifier.wosid000463006000024-
dc.identifier.scopusid2-s2.0-85062468203-
dc.identifier.rimsid67618-
dc.contributor.affiliatedAuthorDae Sik Kim-
dc.contributor.affiliatedAuthorDa-eun Kim-
dc.contributor.affiliatedAuthorGyeorae Lee-
dc.contributor.affiliatedAuthorSung-Ik Cho-
dc.contributor.affiliatedAuthorJin Soo Kim-
dc.identifier.doi10.1038/s41587-019-0050-1-
dc.identifier.bibliographicCitationNATURE BIOTECHNOLOGY, v.37, no.4, pp.430 - 435-
dc.citation.titleNATURE BIOTECHNOLOGY-
dc.citation.volume37-
dc.citation.number4-
dc.citation.startPage430-
dc.citation.endPage435-
dc.description.journalClass1-
dc.description.journalRegisteredClassscie-
dc.description.journalRegisteredClassscopus-
dc.subject.keywordPlusENDONUCLEASES-
dc.subject.keywordPlusDNA-
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > 1. Journal Papers (저널논문)
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