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

Wang, Xiaoling; Wang, Yebo; Wu, Xiwei; Wang, Jinhui; Wang, Yingjia; Qiu, Zhaojun; Chang, Tammy; Huang, He et al. · Nat Biotechnol · 2015

basic_science · Level V

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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.

Medical subject headings