| Chapter title |
time-ChIP: A Method to Determine Long-Term Locus-Specific Nucleosome Inheritance
|
|---|---|
| Chapter number | 7 |
| Book title |
Histone Variants
|
| Published in |
Methods in molecular biology, August 2018
|
| DOI | 10.1007/978-1-4939-8663-7_7 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8662-0, 978-1-4939-8663-7
|
| Authors |
Wojciech Siwek, Mariluz Gómez-Rodríguez, Daniel Sobral, Ivan R. Corrêa, Lars E. T. Jansen |
| Abstract |
Understanding chromatin dynamics is essential to define the contribution of chromatin to heritable gene silencing and the long-term maintenance of gene expression. Here we present a detailed protocol for time-ChIP, a novel method to measure histone turnover at high resolution across long timescales. This method is based on the SNAP-tag, a self-labeling enzyme that can be pulse labeled with small molecules in cells. Upon pulse biotinylation of a cohort of SNAP-tagged histones we can determine their abundance and fate across a chase period using a biotin-specific chromatin pulldown followed by DNA sequencing or quantitative PCR. This method is unique in its ability to trace the long-term fate of a chromatin bound histone pool, genome wide. In addition to a step by step protocol, we outline advantages and limitations of the method in relation to other existing techniques. time-ChIP can define regions of high and low histone turnover and identify the location of pools of long lived histones. |
X Demographics
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|---|---|---|
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| Canada | 1 | 17% |
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| Unknown | 3 | 50% |
Demographic breakdown
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| Scientists | 2 | 33% |
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Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 21 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 6 | 29% |
| Student > Master | 3 | 14% |
| Student > Ph. D. Student | 2 | 10% |
| Student > Bachelor | 1 | 5% |
| Student > Doctoral Student | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 7 | 33% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 8 | 38% |
| Nursing and Health Professions | 2 | 10% |
| Agricultural and Biological Sciences | 2 | 10% |
| Medicine and Dentistry | 1 | 5% |
| Chemistry | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 6 | 29% |