Activating gene expression in mammalian cells with promoter-targeted duplex RNAs

Bethany A. Janowski, Scott T. Younger, Daniel B. Hardy, Rosalyn Ram, Kenneth E. Huffman, David R. Corey

Research output: Contribution to journalArticle

359 Citations (Scopus)

Abstract

The ability to selectively activate or inhibit gene expression is fundamental to understanding complex cellular systems and developing therapeutics. Recent studies have demonstrated that duplex RNAs complementary to promoters within chromosomal DNA are potent gene silencing agents in mammalian cells. Here we report that chromosome-targeted RNAs also activate gene expression. We have identified multiple duplex RNAs complementary to the progesterone receptor (PR) promoter that increase expression of PR protein and RNA after transfection into cultured T47D or MCF7 human breast cancer cells. Upregulation of PR protein reduced expression of the downstream gene encoding cyclooygenase 2 but did not change concentrations of estrogen receptor, which demonstrates that activating RNAs can predictably manipulate physiologically relevant cellular pathways. Activation decreased over time and was sequence specific. Chromatin immunoprecipitation assays indicated that activation is accompanied by reduced acetylation at histones H3K9 and H3K14 and by increased di- and trimethylation at histone H3K4. These data show that, like proteins, hormones and small molecules, small duplex RNAs interact at promoters and can activate or repress gene expression.

Original languageEnglish (US)
Pages (from-to)166-173
Number of pages8
JournalNature Chemical Biology
Volume3
Issue number3
DOIs
StatePublished - Mar 2007

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Progesterone Receptors
RNA
Gene Expression
Complementary RNA
Histones
Proteins
Chromatin Immunoprecipitation
Gene Silencing
Acetylation
Estrogen Receptors
Transfection
Up-Regulation
Chromosomes
Hormones
Breast Neoplasms
DNA
Therapeutics

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Activating gene expression in mammalian cells with promoter-targeted duplex RNAs. / Janowski, Bethany A.; Younger, Scott T.; Hardy, Daniel B.; Ram, Rosalyn; Huffman, Kenneth E.; Corey, David R.

In: Nature Chemical Biology, Vol. 3, No. 3, 03.2007, p. 166-173.

Research output: Contribution to journalArticle

Janowski, Bethany A. ; Younger, Scott T. ; Hardy, Daniel B. ; Ram, Rosalyn ; Huffman, Kenneth E. ; Corey, David R. / Activating gene expression in mammalian cells with promoter-targeted duplex RNAs. In: Nature Chemical Biology. 2007 ; Vol. 3, No. 3. pp. 166-173.
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