Direct observation of DNA target searching and cleavage by CRISPR-Cas12a

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Title
Direct observation of DNA target searching and cleavage by CRISPR-Cas12a
Author(s)
Yongmoon Jeon; You Hee Choi; Yunsu Jang; Jihyeon Yu; Jiyoung Goo; Gyejun Lee; You Kyeong Jeong; Seung Hwan Lee; In-San Kim; Jin-Soo Kim; Cherlhyun Jeong; Sanghwa Lee; Sangsu Bae
Publication Date
2018-07
Journal
NATURE COMMUNICATIONS, v.9, no.1, pp.2777 -
Publisher
NATURE PUBLISHING GROUP
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. © 2018 The Author(s)
URI
https://pr.ibs.re.kr/handle/8788114/5782
ISSN
2041-1723
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
Files in This Item:
성과추가_1807_사사X_김진수_NATURE COMMUNICATIONS.pdfDownload

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