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Science 305 (5688): 1289-1292

Copyright © 2004 by the American Association for the Advancement of Science

Small Interfering RNA-Induced Transcriptional Gene Silencing in Human Cells

Kevin V. Morris,1*{dagger} Simon W.-L. Chan,2 Steven E. Jacobsen,2,3 David J. Looney1,4

Abstract: Small interfering RNA (siRNA) and microRNA silence genes at the transcriptional, posttranscriptional, and/or translational level. Using human tissue culture cells, we show that promoter-directed siRNA inhibits transcription of an integrated, proviral, elongation factor 1alpha (EF1A) promoter–green fluorescent protein reporter gene and of endogenous EF1A. Silencing was associated with DNA methylation of the targeted sequence, and it required either active transport of siRNA into the nucleus or permeabilization of the nuclear envelope by lentiviral transduction. These results demonstrate that siRNA-directed transcriptional silencing is conserved in mammals, providing a means to inhibit mammalian gene function.

1 Department of Medicine 0678, Stein Clinical Research Building, Room 302, University of California–San Diego, La Jolla, CA 92093–0678, USA.
2 Department of Molecular, Cell, and Developmental Biology, University of California–Los Angeles, Los Angeles, CA 90095–1606, USA.
3 Molecular Biology Institute, University of California–Los Angeles, Los Angeles, CA 90095–1606, USA.
4 VA San Diego Healthcare System, Infectious Diseases, San Diego, CA 92161, USA.

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Note added in proof: After this work was submitted, Fukugawa et al. showed that Dicer-defective chicken cells have heterochromatin defects at centromeres, possibly implicating siRNA in centromeric silencing (31).

* Present address: Division of Molecular Biology, Beckman Research Institute, 1450 East Duarte Road, Duarte, CA 91010–3011, USA.

{dagger} To whom correspondence should be addressed. E-mail: kmorris{at}coh.org


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