title: Isolation of specific genomic regions and identification of associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR.
abstract: Isolation of specific genomic regions retaining molecular interactions is necessary for their biochemical analysis. Here, we describe engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using the CRISPR system, for purification of specific genomic regions retaining molecular interactions. In this form of enChIP, specific genomic regions are immunoprecipitated with antibody against a tag(s), which is fused to a catalytically inactive form of Cas9 (dCas9), which is co-expressed with a guide RNA (gRNA) and recognizes endogenous DNA sequence in the genomic regions of interest. enChIP combined with mass spectrometry (enChIP-MS), next-generation sequencing (enChIP-Seq), and RNA-Seq (enChIP-RNA-Seq) can identify proteins, other genomic regions, and RNA, respectively, that interact with the target genomic region.
authors.affiliation: Chromatin Biochemistry Research Group, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita-shi, Osaka, 565-0871, Japan.
authors.forename: Toshitsugu
authors.initials: T
authors.lastname: Fujita
authors.forename: Hodaka
authors.initials: H
authors.lastname: Fujii
mesh_terms: D002843:Chromatin / Q000235:genetics* / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D016678:Genome; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D013058:Mass Spectrometry; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics
chemical_list: D002843:Chromatin; D000094704:RNA, Guide, CRISPR-Cas Systems