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dc.contributor.authorRamakrishna Suresh-
dc.date.accessioned2019-12-06T02:15:50Z-
dc.date.available2019-12-06T02:15:50Z-
dc.date.issued2018-03-
dc.identifier.citationNUCLEIC ACIDS RESEARCH, v. 46, no. 12, Article no. e71en_US
dc.identifier.issn0305-1048-
dc.identifier.issn1362-4962-
dc.identifier.urihttps://academic.oup.com/nar/article/46/12/e71/4951847-
dc.identifier.urihttps://repository.hanyang.ac.kr/handle/20.500.11754/117804-
dc.description.abstractThe use of paired Cas9 nickases instead of Cas9 nuclease drastically reduces off-target effects. Because both nickases must function for a nickase pair to make a double-strand break, the efficiency of paired nickases can intuitively be expected to be lower than that of either corresponding nuclease alone. Here, we carefully compared the gene-disrupting efficiency of Cas9 paired nickases with that of nucleases. Interestingly, the T7E1 assay and deep sequencing showed that on-target efficiency of paired D10A Cas9 nickases was frequently comparable, but sometimes higher than that of either corresponding nucleases in mammalian cells. As the underlying mechanism, we found that the HNH domain, which is preserved in the D10A Cas9 nickase, has higher activity than the RuvC domain in mammalian cells. In this study, we showed: (i) the in vivo cleavage efficiency of the HNH domain of Cas9 in mammalian cells is higher than that of the RuvC domain, (ii) paired Cas9 nickases are sometimes more efficient than individual nucleases for gene disruption. We envision that our findings which were overlooked in previous reports will serve as a new potential guideline for tool selection for CRISPR-Cas9-mediated gene disruption, facilitating efficient and precise genome editing.en_US
dc.description.sponsorshipKorea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea [HI16C1009]; National Research Foundation of Korea [2017M3A9C6061361]. Funding for open access charge: Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea [HI16C1009]; National Research Foundation of Korea [2017M3A9C6061361].en_US
dc.language.isoen_USen_US
dc.publisherOXFORD UNIV PRESSen_US
dc.subjectRNAen_US
dc.subjectSPECIFICITYen_US
dc.subjectCELLSen_US
dc.subjectGUIDEen_US
dc.subjectENRICHMENTen_US
dc.subjectREPAIRen_US
dc.titlePaired D10A Cas9 nickases are sometimes more efficient than individual nucleases for gene disruptionen_US
dc.typeArticleen_US
dc.relation.no12-
dc.relation.volume46-
dc.identifier.doi10.1093/nar/gky222-
dc.relation.page1-12-
dc.relation.journalNUCLEIC ACIDS RESEARCH-
dc.contributor.googleauthorGopalappa, Ramu-
dc.contributor.googleauthorSuresh, Bharathi-
dc.contributor.googleauthorRamakrishna, Suresh-
dc.contributor.googleauthorKim, Hyongbum Henry-
dc.relation.code2018001029-
dc.sector.campusS-
dc.sector.daehakGRADUATE SCHOOL OF BIOMEDICAL SCIENCE AND ENGINEERING[S]-
dc.identifier.pidsuri28-


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