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Nucleic Acids Research 2004 32(21):e171; doi:10.1093/nar/gnh170
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Published online 2 December 2004

Nucleic Acids Research, Vol. 32 No. 21 © Oxford University Press 2004; all rights reserved

RNA silencing in plants by the expression of siRNA duplexes

Shanfa Lu*, Rui Shi, Cheng-Chung Tsao1, Xiaoping Yi, Laigeng Li and Vincent L. Chiang

Forest Biotechnology Group, Department of Forestry, North Carolina State University, Raleigh, NC 27695, USA and 1 Program in Signal Transduction Research, The Burnham Institute, La Jolla, CA 92037, USA

* To whom correspondence should be addressed at Forest Biotechnology Group, Department of Forestry, Campus Box 7247, 2500 Partners II Bldg, 840 Main Campus Drive, North Carolina State University, Raleigh, NC 27695-7247, USA. Tel: +1 919 513 0015; Fax: +1 919 515 7801; Email: slu{at}ncsu.edu

Received August 17, 2004; Accepted November 15, 2004

In animal cells, stable RNA silencing can be achieved by vector-based small interfering RNA (siRNA) expression system, in which Pol III RNA gene promoters are used to drive the expression of short hairpin RNA, however, this has not been demonstrated in plants. Whether Pol III RNA gene promoter is capable of driving siRNA expression in plants is unknown. Here, we report that RNA silencing was achieved in plants through stable expression of short hairpin RNA, which was driven by Pol III RNA gene promoters. Using glucuronidase (GUS) transformed tobacco as a model system, the results demonstrated that 21 nt RNA duplexes, targeting at different sites of GUS gene, were stably expressed under the control of either human H1 or Arabidopsis 7SL RNA gene promoter, and GUS gene was silenced in 80% of siRNA transgenics. The severity of silencing was correlated with the abundance of siRNA expression but independent of the target sites and uridine residue structures in siRNA hairpin transcripts. Thus, the specific expression of siRNA provides a new system for the study of siRNA silencing pathways and functional genomics in plants. Moreover, the effectiveness of the human H1 promoter in a plant background suggested a conserved mechanism underlying Pol III complex functionality.


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