Activating gene expression in mammalian cells with promoter-targeted duplex RNAs.
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.
Janowski
Bethany A
BA
Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390, USA. bethany.janowski@utsouthwestern.edu
Younger
Scott T
ST
Hardy
Daniel B
DB
Ram
Rosalyn
R
Huffman
Kenneth E
KE
Corey
David R
DR
D015153:Blotting, Western; D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism; D003839:Deoxyadenosines / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000502:physiology*; D006657:Histones / Q000235:genetics / Q000502:physiology; D006728:Hormones / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D047468:Immunoprecipitation; D053583:Interleukin-1beta / Q000494:pharmacology; D008969:Molecular Sequence Data; D020135:Peptide Nucleic Acids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012313:RNA / Q000502:physiology*; D012330:RNA, Double-Stranded / Q000235:genetics; D011980:Receptors, Progesterone / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013872:Thionucleosides / Q000494:pharmacology; D015854:Up-Regulation / Q000235:genetics
D002843:Chromatin; D003839:Deoxyadenosines; D004791:Enzyme Inhibitors; D006657:Histones; D006728:Hormones; D006877:Hydroxamic Acids; D053583:Interleukin-1beta; D020135:Peptide Nucleic Acids; D012330:RNA, Double-Stranded; D011980:Receptors, Progesterone; D013872:Thionucleosides; C012589:trichostatin A; D012313:RNA; C008500:5'-methylthioadenosine