Enhancement of target specificity of CRISPR-Cas12a by using a chimeric DNA-RNA guide
DC Field | Value | Language |
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dc.contributor.author | Kim, Hanseop | - |
dc.contributor.author | Lee, Wi-jae | - |
dc.contributor.author | Oh, Yeounsun | - |
dc.contributor.author | Kang, Seung-Hun | - |
dc.contributor.author | Hur, Junho K. | - |
dc.contributor.author | Lee, Hyomin | - |
dc.contributor.author | Song, WooJeung | - |
dc.contributor.author | Lim, Kyung-Seob | - |
dc.contributor.author | Park, Young-Ho | - |
dc.contributor.author | Song, Bong-Seok | - |
dc.contributor.author | Jin, Yeung | - |
dc.contributor.author | Jun, Bong-Hyun | - |
dc.contributor.author | Jung, Cheulhee | - |
dc.contributor.author | Lee, Dong-Seok | - |
dc.contributor.author | Kim, Sun-Uk | - |
dc.contributor.author | Lee, Seung Hwan | - |
dc.date.accessioned | 2021-08-30T14:45:14Z | - |
dc.date.available | 2021-08-30T14:45:14Z | - |
dc.date.created | 2021-06-19 | - |
dc.date.issued | 2020-09-04 | - |
dc.identifier.issn | 0305-1048 | - |
dc.identifier.uri | https://scholar.korea.ac.kr/handle/2021.sw.korea/53188 | - |
dc.description.abstract | The CRISPR-Cas9 system is widely used for target-specific genome engineering. CRISPR-Cas12a (Cpf1) is one of the CRISPR effectors that controls target genes by recognizing thymine-rich protospacer adjacent motif (PAM) sequences. Cas12a has a higher sensitivity to mismatches in the guide RNA than does Cas9; therefore, off-target sequence recognition and cleavage are lower. However, it tolerates mismatches in regions distant from the PAM sequence (TTTN or TTN) in the protospacer, and off-target cleavage issues may become more problematic when Cas12a activity is improved for therapeutic purposes. Therefore, we investigated off-target cleavage by Cas12a and modified the Cas12a (cr)RNA to address the off-target cleavage issue. We developed a CRISPR-Cas12a that can induce mutations in target DNA sequences in a highly specific and effective manner by partially substituting the (cr)RNA with DNA to change the energy potential of base pairing to the target DNA. A model to explain how chimeric (cr)RNA guided CRISPR-Cas12a and SpCas9 nickase effectively work in the intracellular genome is suggested. Chimeric guide-based CRISPR-Cas12a genome editing with reduced off-target cleavage, and the resultant, increased safety has potential for therapeutic applications in incurable diseases caused by genetic mutations. | - |
dc.language | English | - |
dc.language.iso | en | - |
dc.publisher | OXFORD UNIV PRESS | - |
dc.subject | STRUCTURAL BASIS | - |
dc.subject | CPF1 | - |
dc.subject | CAS9 | - |
dc.subject | ENDONUCLEASE | - |
dc.subject | COMPLEX | - |
dc.subject | MICE | - |
dc.title | Enhancement of target specificity of CRISPR-Cas12a by using a chimeric DNA-RNA guide | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Jung, Cheulhee | - |
dc.identifier.doi | 10.1093/nar/gkaa605 | - |
dc.identifier.scopusid | 2-s2.0-85090491150 | - |
dc.identifier.wosid | 000574315100033 | - |
dc.identifier.bibliographicCitation | NUCLEIC ACIDS RESEARCH, v.48, no.15, pp.8601 - 8616 | - |
dc.relation.isPartOf | NUCLEIC ACIDS RESEARCH | - |
dc.citation.title | NUCLEIC ACIDS RESEARCH | - |
dc.citation.volume | 48 | - |
dc.citation.number | 15 | - |
dc.citation.startPage | 8601 | - |
dc.citation.endPage | 8616 | - |
dc.type.rims | ART | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.subject.keywordPlus | STRUCTURAL BASIS | - |
dc.subject.keywordPlus | CPF1 | - |
dc.subject.keywordPlus | CAS9 | - |
dc.subject.keywordPlus | ENDONUCLEASE | - |
dc.subject.keywordPlus | COMPLEX | - |
dc.subject.keywordPlus | MICE | - |
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