期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 439, 期 1, 页码 132-136出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2013.08.013
关键词
enChIP; CRISPR; dCas9; gRNA; Mass spectrometry; Locus-specific biochemical epigenetics/chromatin biochemistry
资金
- Research Institute for Microbial Diseases
- Program for Combined Research Fields from Immunology Frontier Research Center
- Osaka University
- Takeda Science Foundation
- Asahi Glass Foundation
- Ministry of Education, Culture, Sports, Science and Technology of Japan [25830131, 23118516, 23114707, 25118512]
- Grants-in-Aid for Scientific Research [25830131, 25118512] Funding Source: KAKEN
Isolation of specific genomic regions retaining molecular interactions is necessary for their biochemical analysis. Here, we established a novel method, engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP), for purification of specific genomic regions retaining molecular interactions. We showed that enChIP using the CRISPR system efficiently isolates specific genomic regions. 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-mass spectrometry (enChIP-MS) targeting endogenous loci identified associated proteins. enChIP using the CRISPR system would be a convenient and useful tool for dissecting chromatin structure of genomic regions of interest. (C) 2013 The Authors. Published by Elsevier Inc. All rights reserved.
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