Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.author | 배상수 | - |
dc.date.accessioned | 2019-12-08T13:56:10Z | - |
dc.date.available | 2019-12-08T13:56:10Z | - |
dc.date.issued | 2018-07 | - |
dc.identifier.citation | NATURE COMMUNICATIONS, v. 9, Article no. 2777 | en_US |
dc.identifier.issn | 2041-1723 | - |
dc.identifier.uri | https://www.nature.com/articles/s41467-018-05245-x | - |
dc.identifier.uri | https://repository.hanyang.ac.kr/handle/20.500.11754/119350 | - |
dc.description.abstract | Cas12a (also called Cpf1) is a representative type V-A CRISPR effector RNA-guided DNA endonuclease, which provides an alternative to type II CRISPR-Cas9 for genome editing. Previous studies have revealed that Cas12a has unique features distinct from Cas9, but the detailed mechanisms of target searching and DNA cleavage by Cas12a are still unclear. Here, we directly observe this entire process by using single-molecule fluorescence assays to study Cas12a from Acidaminococcus sp. (AsCas12a). We determine that AsCas12a ribonucleoproteins search for their on-target site by a one-dimensional diffusion along elongated DNA molecules and induce cleavage in the two DNA strands in a well-defined order, beginning with the non-target strand. Furthermore, the protospacer-adjacent motif (PAM) for AsCas12a makes only a limited contribution of DNA unwinding during R-loop formation and shows a negligible role in the process of DNA cleavage, in contrast to the Cas9 PAM. | en_US |
dc.description.sponsorship | We thank Prof. J.-B. Lee (POSTECH) for the helpful advice and discussion. This work was supported by National Research Foundation of Korea (NRF) Grants (No. 2017R1A6A3A01007932 to Yo.J.; 2016R1A6A3A11930747 to Y.H.C.; 2017R1A3B1023418 and 2018R1A2B2008995 to C.J.; 2015R1C1A1A01055164 and 2016R1A5A1007318 to S.L.; 2017M3A9G8084539 and 2018M3A9H3022412 to S.B.), and by a grant from the KIST Institutional Program to C.J., and by a grant from the National R&D Program for Cancer Control of Ministry (1520090), the GIST Research Insitute to S.L., and by grants from Next Generation BioGreen 21 Program (PJ01319301), Korea Healthcare technology R&D Project (HI16C1012) to S.B. | en_US |
dc.language.iso | en_US | en_US |
dc.publisher | NATURE PUBLISHING GROUP | en_US |
dc.subject | RNA-GUIDED ENDONUCLEASE | en_US |
dc.subject | REAL-TIME OBSERVATION | en_US |
dc.subject | CRISPR-CAS SYSTEM | en_US |
dc.subject | R-LOOP FORMATION | en_US |
dc.subject | STRUCTURAL BASIS | en_US |
dc.subject | CRYSTAL-STRUCTURE | en_US |
dc.subject | MISMATCH REPAIR | en_US |
dc.subject | HUMAN-CELLS | en_US |
dc.subject | CPF1 | en_US |
dc.subject | COMPLEX | en_US |
dc.title | Direct observation of DNA target searching and cleavage by CRISPR-Cas12a | en_US |
dc.type | Article | en_US |
dc.relation.volume | 9 | - |
dc.identifier.doi | 10.1038/s41467-018-05245-x | - |
dc.relation.page | 1-11 | - |
dc.relation.journal | NATURE COMMUNICATIONS | - |
dc.contributor.googleauthor | Jeon, Yongmoon | - |
dc.contributor.googleauthor | Choi, You Hee | - |
dc.contributor.googleauthor | Jang, Yunsu | - |
dc.contributor.googleauthor | Yu, Jihyeon | - |
dc.contributor.googleauthor | Goo, Jiyoung | - |
dc.contributor.googleauthor | Lee, Gyejun | - |
dc.contributor.googleauthor | Jeong, You Kyeong | - |
dc.contributor.googleauthor | Lee, Seung Hwan | - |
dc.contributor.googleauthor | Kim, In-San | - |
dc.contributor.googleauthor | Bae, Sangsu | - |
dc.relation.code | 2018003595 | - |
dc.sector.campus | S | - |
dc.sector.daehak | COLLEGE OF NATURAL SCIENCES[S] | - |
dc.sector.department | DEPARTMENT OF CHEMISTRY | - |
dc.identifier.pid | sangsubae | - |
dc.identifier.researcherID | E-5324-2017 | - |
dc.identifier.orcid | http://orcid.org/0000-0003-3615-8566 | - |
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