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PNAS 103 (46): 17337-17342

Copyright © 2006 by the National Academy of Sciences.

BIOLOGICAL SCIENCES / GENETICS

Small dsRNAs induce transcriptional activation in human cells

Long-Cheng Li*,, Steven T. Okino, Hong Zhao, Deepa Pookot, Robert F. Place, Shinji Urakami, Hideki Enokida, and Rajvir Dahiya*,

Department of Urology, Veterans Affairs Medical Center and University of California, San Francisco, CA 94121

Edited by Mark T. Groudine, Fred Hutchinson Cancer Research Center, Seattle, WA, and approved September 28, 2006

Received for publication August 15, 2006.

Abstract: Recent studies have shown that small noncoding RNAs, such as microRNAs and siRNAs, regulate gene expression at multiple levels including chromatin architecture, transcription, RNA editing, RNA stability, and translation. Each form of RNA-dependent regulation has been generally found to silence homologous sequences and collectively called RNAi. To further study the regulatory role of small RNAs at the transcriptional level, we designed and synthesized 21-nt dsRNAs targeting selected promoter regions of human genes E-cadherin, p21WAF1/CIP1 (p21), and VEGF. Surprisingly, transfection of these dsRNAs into human cell lines caused long-lasting and sequence-specific induction of targeted genes. dsRNA mutation studies reveal that the 5' end of the antisense strand, or "seed" sequence, is critical for activity. Mechanistically, the dsRNA-induced gene activation requires the Argonaute 2 (Ago2) protein and is associated with a loss of lysine-9 methylation on histone 3 at dsRNA-target sites. In conclusion, we have identified several dsRNAs that activate gene expression by targeting noncoding regulatory regions in gene promoters. These findings reveal a more diverse role for small RNA molecules in the regulation of gene expression than previously recognized and identify a potential therapeutic use for dsRNA in targeted gene activation.

Key Words: gene regulation • promoter • Argonaute 2

Author contributions: L.-C.L. designed research; L.-C.L., S.T.O., H.Z., D.P., R.F.P., S.U., and H.E. performed research; L.-C.L. and R.F.P. analyzed data; and L.-C.L., R.F.P., and R.D. wrote the paper.

The authors declare no conflict of interest.

This article is a PNAS direct submission.

*To whom correspondence may be addressed at: Urology Research Center, Veterans Affairs Medical Center and University of California, 4150 Clement Street, San Francisco, CA 94121. E-mail: longcheng.li{at}ucsf.edu or rdahiya{at}urology.ucsf.edu

© 2006 by The National Academy of Sciences of the USA

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