Detecting and mitigating off-target activity is critical to the practical application of CRISPR-mediated genome and epigenome editing. While numerous methods have been developed to map genome-wide sgRNA specificity genome-wide, they are generally time-consuming and/or expensive, and not applicable to catalytically dead CRISPR enzymes. We have developed a rapid, inexpensive, and facile assay for identifying off-target CRISPR binding and cleavage by chemically mapping the unwound single-stranded DNA structures formed upon CRISPR binding (``CasKAS''). We demonstrate this method in both in vitro and in vivo contexts.
Overall design
We have developed a rapid, inexpensive, and facile assay for identifying off-target CRISPR binding and cleavage by chemically mapping the unwound single-stranded DNA structures formed upon CRISPR binding (``CasKAS'').
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