Hunter, Ann Louise;
Adamson, Antony;
Poolman, Toryn;
Grudzien, Magdalena;
Loudon, Andrew;
Ray, David;
Bechtold, David A;
(2022)
HaloChIP-seq for Antibody-Independent Mapping of Mouse Transcription Factor Cistromes In Vivo.
bio-protocol
, 12
(13)
, Article e4460. 10.21769/bioprotoc.4460.
Preview |
Text
Bio-protocol4460.pdf - Other Download (1MB) | Preview |
Abstract
Chromatin immunoprecipitation (ChIP) maps, on a genome-wide scale, transcription factor binding sites, and the distribution of other chromatin-associated proteins and their modifications. As such, it provides valuable insights into mechanisms of gene regulation. However, successful ChIP experiments are dependent on the availability of a high-quality antibody against the target of interest. Using antibodies with poor sensitivity and specificity can yield misleading results. This can be partly circumvented by using epitope-tagged systems (e.g., HA, Myc, His), but these approaches are still antibody-dependent. HaloTag® is a modified dehalogenase enzyme, which covalently binds synthetic ligands. This system can be used for imaging and purification of HaloTag ® fusion proteins, and has been used for ChIP in vitro. Here, we present a protocol for using the HaloTag® system for ChIP in vivo, to map, with sensitivity and specificity, the cistrome of a dynamic mouse transcription factor expressed at its endogenous locus.
| Type: | Article |
|---|---|
| Title: | HaloChIP-seq for Antibody-Independent Mapping of Mouse Transcription Factor Cistromes In Vivo |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.21769/bioprotoc.4460 |
| Publisher version: | https://doi.org/10.21769/bioprotoc.4460 |
| Language: | English |
| Additional information: | © Hunter et al. This article is distributed under the terms of the Creative Commons Attribution License CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/). |
| Keywords: | HaloTag®, ChIP, Nuclear receptor, Tag, Fusion protein, NR1D1 |
| UCL classification: | UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10151993 |
Archive Staff Only
 |
View Item |
