Profiling genome-wide specificity of CRISPR-Cas9 using digenome-seq

Daesik Kim, Jin Soo Kim

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Scopus citations

Abstract

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages233-242
Number of pages10
DOIs
StatePublished - 2021
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2162
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cas9
  • CRISPR
  • Genome editing
  • Off-target
  • Whole-genome sequencing

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