Rapid and reversible epigenome editing by endogenous chromatin regulators
| Autor: | Dylan Husmann, Gerald R. Crabtree, Jacob G. Kirkland, Simon M. G. Braun, Joseph P Calarco, Emma J. Chory |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Science Polycomb-Group Proteins General Physics and Astronomy Polycomb-Group Proteins/metabolism Heterochromatin/metabolism Biology Article General Biochemistry Genetics and Molecular Biology Chromatin remodeling Epigenesis Genetic Chromatin/metabolism Promoter Regions 03 medical and health sciences Genetic Heterochromatin Epigenome editing Humans Epigenetics Gene Silencing Promoter Regions Genetic lcsh:Science ChIA-PET Epigenomics Genetics Gene Editing Multidisciplinary General Chemistry Epigenome Chromatin Assembly and Disassembly Chromatin Cell biology 030104 developmental biology HEK293 Cells Gene Expression Regulation lcsh:Q CRISPR-Cas Systems Bivalent chromatin Epigenesis |
| Zdroj: | Nature Communications, Vol. 8, No 1 (2017) P. 560 Nature Communications, Vol 8, Iss 1, Pp 1-8 (2017) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | Understanding the causal link between epigenetic marks and gene regulation remains a central question in chromatin biology. To edit the epigenome we developed the FIRE-Cas9 system for rapid and reversible recruitment of endogenous chromatin regulators to specific genomic loci. We enhanced the dCas9–MS2 anchor for genome targeting with Fkbp/Frb dimerizing fusion proteins to allow chemical-induced proximity of a desired chromatin regulator. We find that mSWI/SNF (BAF) complex recruitment is sufficient to oppose Polycomb within minutes, leading to activation of bivalent gene transcription in mouse embryonic stem cells. Furthermore, Hp1/Suv39h1 heterochromatin complex recruitment to active promoters deposits H3K9me3 domains, resulting in gene silencing that can be reversed upon washout of the chemical dimerizer. This inducible recruitment strategy provides precise kinetic information to model epigenetic memory and plasticity. It is broadly applicable to mechanistic studies of chromatin in mammalian cells and is particularly suited to the analysis of endogenous multi-subunit chromatin regulator complexes. Understanding the link between epigenetic marks and gene regulation requires the development of new tools to directly manipulate chromatin. Here the authors demonstrate a Cas9-based system to recruit chromatin remodelers to loci of interest, allowing rapid, reversible manipulation of epigenetic states. |
| Databáze: | OpenAIRE |
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