Genome-wide target specificities of CRISPR RNA-guided programmable deaminases Highly Cited Paper

Cited 29 time in webofscience Cited 0 time in scopus
310 Viewed 228 Downloaded
Title
Genome-wide target specificities of CRISPR RNA-guided programmable deaminases
Author(s)
Daesik Kim; Kayeong Lim; Sang-Tae Kim; Sun-heui Yoon; Kyoungmi Kim; Seuk-Min Ryu; Jin-Soo Kim
Publication Date
2017-05
Journal
NATURE BIOTECHNOLOGY, v.35, no.5, pp.475 - 480
Publisher
NATURE PUBLISHING GROUP
Abstract
Cas9-linked deaminases, also called base editors, enable targeted mutation of single nucleotides in eukaryotic genomes. However, their off-target activity is largely unknown. Here we modify digested-genome sequencing (Digenome-seq) to assess the specificity of a programmable deaminase composed of a Cas9 nickase (nCas9) and the deaminase APOBEC1 in the human genome. Genomic DNA is treated with the base editor and a mixture of DNA-modifying enzymes in vitro to produce DNA double-strand breaks (DSBs) at uracil-containing sites. Off-target sites are then computationally identified from whole genome sequencing data. Testing seven different single guide RNAs (sgRNAs), we find that the rAPOBEC1-nCas9 base editor is highly specific, inducing cytosine-to-uracil conversions at only 18 ± 9 sites in the human genome for each sgRNA. Digenome-seq is sensitive enough to capture off-target sites with a substitution frequency of 0.1%. Notably, off-target sites of the base editors are often different from those of Cas9 alone, calling for independent assessment of their genome-wide specificities. © 2017 Nature America, Inc., part of Springer Nature. All rights reserved.
URI
https://pr.ibs.re.kr/handle/8788114/3703
ISSN
1087-0156
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
Files in This Item:
김대식_nbt.3852.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