Elucidation of molecular mechanisms of genome functions such as transcription and epigenetic regulation requires identification of components mediating the genome functions. To this end, we recently developed the locus-specific chromatin immunoprecipitation (ChIP) (locus-specific ChIP) technologies to identify molecules interacting with a given genomic region of interest in vivo.
Locus-specific ChIP consists of insertional ChIP (iChIP) and engineered DNA-binding molecule-mediated ChIP (enChIP) using transcription activator-like (TAL) proteins and the clustered regularly interspaced short palindromic repeats (CRISPR) system. Basically, locus-specific ChIP consists of locus tagging and affinity purification and can be combined with down-stream analyses such as mass spectrometry (MS) (iChIP-MS and enChIP-MS) and RNA sequencing (RNA-Seq) (iChIP-RNA-Seq and enChIP-RNA-Seq) to identify proteins and RNAs associated with the target genomic region.
Among others, enChIP using CRISPR is the most flexible and easy-to-use method in locus-specific ChIP because any genomic regions of interest can be easily targeted with guide RNAs. In this webinar, an overview of locus-specific ChIP will be given. In addition, applications of locus-specific ChIP will be introduced. Furthermore, step-by-step protocols of enChIP using CRISPR will be presented.