Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors
Nature biotechnology, 2015•nature.com
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.
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.
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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