Analysis of Cas9 binding specificity in Saccharomyces cerevisiae

Utilizing the CRISPR system, we previously developed a methodology for enriching a specific genomic locus of interest for proteomic analysis in Saccharomyces cerevisiae. This approach utilized a guide RNA-targeted, catalytically dead Cas9 (dCas9) as an affinity reagent. To more comprehensively evaluate the genomic specificity of this technique, we performed dCas9-mediated locus enrichment followed by next-generation sequencing on a genome-wide scale.