BROWSE

Related Scientist

Researcher

나노의학 연구단
나노의학 연구단
more info

High-throughput analysis of the activities of xCas9, SpCas9-NG and SpCas9 at matched and mismatched target sequences in human cells

Cited 0 time in webofscience Cited 0 time in scopus
39 Viewed 9 Downloaded
Title
High-throughput analysis of the activities of xCas9, SpCas9-NG and SpCas9 at matched and mismatched target sequences in human cells
Author(s)
Hui Kwon Kim; Sungtae Lee; Younggwang Kim; Jinman Park; Seonwoo Min; Jae Woo Choi; Tony P. Huang; Sungroh Yoon; David R. Liu; Hyongbum Henry Kim
Publication Date
2020-01
Journal
NATURE BIOMEDICAL ENGINEERING, v.4, no.1, pp.111 - 124
Publisher
NATURE PUBLISHING GROUP
Abstract
© 2020, The Author(s), under exclusive licence to Springer Nature Limited.The applications of clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing can be limited by a lack of compatible protospacer adjacent motifs (PAMs), insufficient on-target activity and off-target effects. Here, we report an extensive comparison of the PAM-sequence compatibilities and the on-target and off-target activities of Cas9 from Streptococcus pyogenes (SpCas9) and the SpCas9 variants xCas9 and SpCas9-NG (which are known to have broader PAM compatibility than SpCas9) at 26,478 lentivirally integrated target sequences and 78 endogenous target sites in human cells. We found that xCas9 has the lowest tolerance for mismatched target sequences and that SpCas9-NG has the broadest PAM compatibility. We also show, on the basis of newly identified non-NGG PAM sequences, that SpCas9-NG and SpCas9 can edit six previously unedited endogenous sites associated with genetic diseases. Moreover, we provide deep-learning models that predict the activities of xCas9 and SpCas9-NG at the target sequences. The resulting deeper understanding of the activities of xCas9, SpCas9-NG and SpCas9 in human cells should facilitate their use
URI
https://pr.ibs.re.kr/handle/8788114/7188
ISSN
2157-846X
Appears in Collections:
Center for Nanomedicine (나노의학 연구단) > Journal Papers
Files in This Item:
202001_High-throughput analysis of the activities of xCas9, SpCas9-NG and SpCas9 at matched and mismatched target sequences in human cells.pdfDownload

qrcode

  • facebook

    twitter

  • Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.
해당 아이템을 이메일로 공유하기 원하시면 인증을 거치시기 바랍니다.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse