HaloChIP-seq for Antibody-Independent Mapping of Mouse Transcription Factor Cistromes in vivo.
Bio Protoc
; 12(13)2022 Jul 05.
Article
en En
| MEDLINE
| ID: mdl-35937930
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. Graphical abstract.
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Colección:
01-internacional
Base de datos:
MEDLINE
Idioma:
En
Revista:
Bio Protoc
Año:
2022
Tipo del documento:
Article
País de afiliación:
Reino Unido