Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells
DC Field | Value | Language |
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dc.contributor.author | Kim, Daesik | - |
dc.contributor.author | Kim, Jungeun | - |
dc.contributor.author | Hur, Jun ho | - |
dc.contributor.author | Been, Kyung Wook | - |
dc.contributor.author | Yoon, Sun-heui | - |
dc.contributor.author | Kim, Jin-Soo | - |
dc.date.accessioned | 2022-07-15T09:48:15Z | - |
dc.date.available | 2022-07-15T09:48:15Z | - |
dc.date.created | 2021-05-14 | - |
dc.date.issued | 2016-08 | - |
dc.identifier.issn | 1087-0156 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/154122 | - |
dc.description.abstract | Programmable clustered regularly interspaced short palindromic repeats (CRISPR) Cpf1 endonucleases are single-RNA-guided (crRNA) enzymes that recognize thymidine-rich protospacer-adjacent motif (PAM) sequences and produce cohesive double-stranded breaks (DSBs). Genome editing with CRISPR-Cpf1 endonucleases could provide an alternative to CRISPR-Cas9 endonucleases, but the determinants of targeting specificity are not well understood. Using mismatched crRNAs we found that Cpf1 could tolerate single or double mismatches in the 3′ PAM-distal region, but not in the 5′ PAM-proximal region. Genome-wide analysis of cleavage sites in vitro for eight Cpf1 nucleases using Digenome-seq revealed that there were 6 (LbCpf1) and 12 (AsCpf1) cleavage sites per crRNA in the human genome, fewer than are present for Cas9 nucleases (>90). Most Cpf1 off-target cleavage sites did not produce mutations in cells. We found mismatches in either the 3′ PAM-distal region or in the PAM sequence of 12 off-target sites that were validated in vivo. Off-target effects were completely abrogated by using preassembled, recombinant Cpf1 ribonucleoproteins. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | NATURE PUBLISHING GROUP | - |
dc.title | Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Hur, Jun ho | - |
dc.identifier.doi | 10.1038/nbt.3609 | - |
dc.identifier.scopusid | 2-s2.0-84981318543 | - |
dc.identifier.wosid | 000381339600024 | - |
dc.identifier.bibliographicCitation | NATURE BIOTECHNOLOGY, v.34, no.8, pp.863 - 868 | - |
dc.relation.isPartOf | NATURE BIOTECHNOLOGY | - |
dc.citation.title | NATURE BIOTECHNOLOGY | - |
dc.citation.volume | 34 | - |
dc.citation.number | 8 | - |
dc.citation.startPage | 863 | - |
dc.citation.endPage | 868 | - |
dc.type.rims | ART | - |
dc.type.docType | 정기학술지(Article(Perspective Article포함)) | - |
dc.description.journalClass | 1 | - |
dc.description.isOpenAccess | N | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biotechnology & Applied Microbiology | - |
dc.subject.keywordPlus | Cpf1 protein | - |
dc.subject.keywordPlus | DNA methyltransferase 1 | - |
dc.subject.keywordPlus | endonuclease | - |
dc.subject.keywordPlus | genomic DNA | - |
dc.subject.keywordPlus | guide RNA | - |
dc.subject.keywordPlus | hypoxanthine phosphoribosyltransferase | - |
dc.subject.keywordPlus | nucleotide | - |
dc.subject.keywordPlus | oligonucleotide | - |
dc.subject.keywordPlus | protein | - |
dc.subject.keywordPlus | ribonucleoprotein | - |
dc.subject.keywordPlus | unclassified drug | - |
dc.subject.keywordPlus | bacterial protein | - |
dc.subject.keywordPlus | endonuclease | - |
dc.subject.keywordPlus | protein binding | - |
dc.identifier.url | https://www.nature.com/articles/nbt.3609 | - |
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