Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 25599175.
- Also identified by DOI 10.1038/nbt.3127.
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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
- CRISPR-Cas Systems
- Lentivirus
- RNA Editing