Synthetic reversal of epigenetic silencing

Karmella Haynes, Pamela A. Silver

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Controlling cell fate-determining gene expression is key to stem cell differentiation, tissue regeneration, and cancer therapy. To date, custom-built transcription factors recognize the information encoded in specific DNA sequences. Chromatin proteins undergo covalent modifications and form complexes that encode a second layer of information that determines proximal gene activity. Here, we employ a novel gene-targeting approach that exploits a specific chromatin modification to reactivate silenced loci in human cells. We used the human Polycomb chromatin protein and homologues from other species to construct modular synthetic transcription factors, called Pc-TFs, that recognize the repressive trimethyl-histone H3 lysine 27 (H3K27me3) signal and switch silenced genes to an active state. Pc-TF expression in U2OS osteosarcoma cells leads to increased transcription of the senescence locus CDKN2A (p16) and other loci in a chromodomain- and activation module-dependent manner, a switch to a senescence phenotype, and reduced cell proliferation. These results indicate that silenced developmental regulators can be reactivated by a synthetic transcription factor that interacts with chromatin rather than DNA, resulting in an altered cell state. As such, our work extends the flexibility of transcription factor engineering and is the first example of chromatin-mediated synthetic transcription factor targeting.

Original languageEnglish (US)
Pages (from-to)27176-27182
Number of pages7
JournalJournal of Biological Chemistry
Volume286
Issue number31
DOIs
StatePublished - Aug 5 2011
Externally publishedYes

Fingerprint

Epigenomics
Chromatin
Transcription Factors
Genes
Switches
Switch Genes
Tissue regeneration
Gene Targeting
DNA sequences
Cell proliferation
Osteosarcoma
Transcription
Stem cells
Gene expression
Histones
Lysine
Regeneration
Cell Differentiation
Proteins
Stem Cells

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Synthetic reversal of epigenetic silencing. / Haynes, Karmella; Silver, Pamela A.

In: Journal of Biological Chemistry, Vol. 286, No. 31, 05.08.2011, p. 27176-27182.

Research output: Contribution to journalArticle

Haynes, Karmella ; Silver, Pamela A. / Synthetic reversal of epigenetic silencing. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 31. pp. 27176-27182.
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