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J Vis Exp. 2019 Aug 23;(150). doi: 10.3791/60039.

A Method to Study de novo Formation of Chromatin Domains.

Author information

1
Howard Hughes Medical Institute, New York University School of Medicine; Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine; Whitehead Institute for Biomedical Research; oksuzo@wi.mit.edu.
2
Howard Hughes Medical Institute, New York University School of Medicine; Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine.

Abstract

The organization and structure of chromatin domains are unique to individual cell lineages. Their misregulation might lead to a loss in cellular identity and/or disease. Despite tremendous efforts, our understanding of the formation and propagation of chromatin domains is still limited. Chromatin domains have been studied under steady-state conditions, which are not conducive to following the initial events during their establishment. Here, we present a method to inducibly reconstruct chromatin domains and follow their re-formation as a function of time. Although, first applied to the case of PRC2-mediated repressive chromatin domain formation, it could be easily adapted to other chromatin domains. The modification of and/or the combination of this method with genomics and imaging technologies will provide invaluable tools to study the establishment of chromatin domains in great detail. We believe that this method will revolutionize our understanding of how chromatin domains form and interact with each other.

PMID:
31498316
DOI:
10.3791/60039

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