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Genome-wide target specificities of CRISPR RNA-guided programmable deaminases

Cited 110 time in webofscience Cited 114 time in scopus
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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
DOI
10.1038/nbt.3852
ISSN
1087-0156
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > 1. Journal Papers (저널논문)
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