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연구성과 > 금주의 우수논문

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SCI Article

Direct observation of DNA target searching and cleavage by CRISPR-Cas12a
성명 배상수()
소속 자연과학대학 화학과
캠퍼스
우수선정주 2018년 08월 2째주
Author Yu, Jihyeon (Dept Chem); Jeong, You Kyeong (Dept Chem); 배상수 (Dept Chem); Yu, Jihyeon (Res Inst Convergence Basic Sci); Bae, Sangsu (Res Inst Convergence Basic Sci);
Corresponding Author Info Jeong, C (reprint author), Korea Inst Sci & Technol, Ctr Theragnosis, Seoul 02792, South Korea.; Lee, S (reprint author), Gwangju Inst Sci & Technol, Adv Photon Res Inst, Gwangju 61005, South Korea.; Lee, S (reprint author), Gwangju Inst Sci & Technol, Ce
E-mail 이메일 아이콘sangsubae@hanyang.ac.kr
Document Type Article
Source NATURE COMMUNICATIONS Volume:9 Issue: Pages:- Published:2018
Times Cited 0
External Information http://dx.doi.org/10.1038/s41467-018-05245-x
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.
Web of Science Categories Multidisciplinary Sciences
Funding National Research Foundation of Korea (NRF) [2017R1A6A3A01007932, 2016R1A6A3A11930747, 2017R1A3B1023418, 2018R1A2B2008995, 2015R1C1A1A01055164, 2016R1A5A1007318, 2017M3A9G8084539, 2018M3A9H3022412]; KIST Institutional Program; National R&D Program for Can
Language English
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