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

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dc.contributor.authorKim, Daesik-
dc.contributor.authorKim, Da-eun-
dc.contributor.authorLee, Gyeorae-
dc.contributor.authorCho, Sung-Ik-
dc.contributor.authorKim, Jin-Soo-
dc.date.accessioned2020-04-27T13:02:33Z-
dc.date.available2020-04-27T13:02:33Z-
dc.date.issued2019-04-
dc.identifier.citationNature Biotechnology, Vol.37 No.4, pp.430-435-
dc.identifier.issn1087-0156-
dc.identifier.other95257-
dc.identifier.urihttps://hdl.handle.net/10371/165712-
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 sequencings(5-8), to assess genomewide 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 RNAs5,8-11, and Sniper/Cas9 (ref.(12)) reduces adenine base editor off-target activity in human cells.-
dc.titleGenome-wide target specificity of CRISPR RNA-guided adenine base editors-
dc.typeArticle-
dc.contributor.AlternativeAuthor김진수-
dc.identifier.doi10.1038/s41587-019-0050-1-
dc.citation.journaltitleNature Biotechnology-
dc.identifier.scopusid2-s2.0-85062468203-
dc.citation.endpage435-
dc.citation.number4-
dc.citation.startpage430-
dc.citation.volume37-
dc.identifier.rimsid95257-
dc.identifier.sci000463006000024-
dc.description.isOpenAccessN-
dc.contributor.affiliatedAuthorKim, Jin-Soo-
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