Digenome-seq is a highly sensitive method for analyzing the genome-wide specificity of CRISPR-Cas9 nuclease activity. In this procedure, genomic DNA is first subjected to digestion by CRISPR-Cas9 in vitro and then to whole genome sequencing, which results in unusual patterns of straight alignments at on-target and potential off-target sites. Analysis of these data with the Digenome-seq computer program allows for identification of the in vitro cleavage sites associated with the straight alignments. Here, we present a detailed Digenome-seq protocol for genome-wide profiling of in vitro CRISPR-Cas9 nuclease cleavage sites.
CITATION STYLE
Kim, D., & Kim, J. S. (2021). Profiling genome-wide specificity of CRISPR-Cas9 using digenome-seq. In Methods in Molecular Biology (Vol. 2162, pp. 233–242). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0687-2_13
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