| Autor: |
Wojciech, Siwek, Mariluz, Gómez-Rodríguez, Daniel, Sobral, Ivan R, Corrêa, Lars E T, Jansen |
| Rok vydání: |
2018 |
| Předmět: |
|
| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 1832 |
| ISSN: |
1940-6029 |
| Popis: |
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. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
