Repressing gene transcription by redirecting cellular machinery with chemical epigenetic modifiers

Anna M. Chiarella, Tiffany A. Wang, Kyle V. Butler, Jian Jin, Nathaniel A. Hathaway

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Regulation of chromatin compaction is an important process that governs gene expression in higher eukaryotes. Although chromatin compaction and gene expression regulation are commonly disrupted in many diseases, a locus-specific, endogenous, and reversible method to study and control these mechanisms of action has been lacking. To address this issue, we have developed and characterized novel gene-regulating bifunctional molecules. One component of the bifunctional molecule binds to a DNA-protein anchor so that it will be recruited to an allele-specific locus. The other component engages endogenous cellular chromatin-modifying machinery, recruiting these proteins to a gene of interest. These small molecules, called chemical epigenetic modifiers (CEMs), are capable of controlling gene expression and the chromatin environment in a dose-dependent and reversible manner. Here, we detail a CEM approach and its application to decrease gene expression and histone tail acetylation at a Green Fluorescent Protein (GFP) reporter located at the Oct4 locus in mouse embryonic stem cells (mESCs). We characterize the lead CEM (CEM23) using fluorescent microscopy, flow cytometry, and chromatin immunoprecipitation (ChIP), followed by a quantitative polymerase chain reaction (qPCR). While the power of this system is demonstrated at the Oct4 locus, conceptually, the CEM technology is modular and can be applied in other cell types and at other genomic loci.

Original languageEnglish
Article numbere58222
JournalJournal of Visualized Experiments
Volume2018
Issue number139
DOIs
StatePublished - 20 Sep 2018

Keywords

  • Bifunctional molecules
  • Bioengineering
  • CEMs
  • Chemical biology
  • Chemical epigenetic modifiers
  • Chemical induced proximity
  • Chromatin regulation
  • Epigenetics
  • Gene repression
  • Histone deacetylase
  • Issue 139

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