0 96

Cited 4 times in

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

Authors
 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 
Citation
 NATURE BIOMEDICAL ENGINEERING, Vol.4(1) : 111-124, 2020 
Journal Title
 NATURE BIOMEDICAL ENGINEERING 
Issue Date
2020
Abstract
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.
Full Text
https://www.nature.com/articles/s41551-019-0505-1
DOI
10.1038/s41551-019-0505-1
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
Yonsei Authors
Kim, Younggwang(김영광) ORCID logo https://orcid.org/0000-0002-8033-4232
Kim, Hyongbum(김형범) ORCID logo https://orcid.org/0000-0002-4693-738X
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/175503
사서에게 알리기
  feedback

qrcode

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

Browse

Links