| 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. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 2 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 3 | 1% |
| United Kingdom | 1 | <1% |
| Argentina | 1 | <1% |
| Brazil | 1 | <1% |
| Unknown | 219 | 97% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 63 | 28% |
| Researcher | 48 | 21% |
| Student > Master | 23 | 10% |
| Student > Bachelor | 23 | 10% |
| Student > Postgraduate | 11 | 5% |
| Other | 33 | 15% |
| Unknown | 24 | 11% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 88 | 39% |
| Agricultural and Biological Sciences | 83 | 37% |
| Chemistry | 7 | 3% |
| Medicine and Dentistry | 5 | 2% |
| Immunology and Microbiology | 4 | 2% |
| Other | 12 | 5% |
| Unknown | 26 | 12% |