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. 2015 Feb;33(2):175-8.
doi: 10.1038/nbt.3127. Epub 2015 Jan 19.

Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors

Xiaoling Wang  1 , Yebo Wang  2 , Xiwei Wu  3 , Jinhui Wang  3 , Yingjia Wang  2 , Zhaojun Qiu  3 , Tammy Chang  1 , He Huang  4 , Ren-Jang Lin  3 , Jiing-Kuan Yee  1
Affiliations

Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors

Xiaoling Wang et al. Nat Biotechnol. 2015 Feb.

Abstract

The utility of CRISPR-Cas9 and TALENs for genome editing may be compromised by their off-target activity. We show that integrase-defective lentiviral vectors (IDLVs) can detect such off-target cleavage with a frequency as low as 1%. In the case of Cas9, we find frequent off-target sites with a one-base bulge or up to 13 mismatches between the single guide RNA (sgRNA) and its genomic target, which refines sgRNA design.

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Comment in

  • Mapping the precision of genome editing.
    Gabriel R, von Kalle C, Schmidt M. Gabriel R, et al. Nat Biotechnol. 2015 Feb;33(2):150-2. doi: 10.1038/nbt.3142. Nat Biotechnol. 2015. PMID: 25658281 No abstract available.

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