Chapter title |
Isolation of Specific Genomic Regions and Identification of Associated Molecules by Engineered DNA-Binding Molecule-Mediated Chromatin Immunoprecipitation (enChIP) Using CRISPR.
|
---|---|
Chapter number | 4 |
Book title |
Chromatin Protocols
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2474-5_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2473-8, 978-1-4939-2474-5
|
Authors |
Fujita, Toshitsugu, Fujii, Hodaka, Toshitsugu Fujita, Hodaka Fujii |
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. |
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Mendeley readers
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