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First published online June 9, 2006; 10.1104/pp.106.082271

Plant Physiology 141:1508-1518 (2006)
© 2006 American Society of Plant Biologists

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SYSTEMS BIOLOGY, MOLECULAR BIOLOGY, AND GENE REGULATION

New Construct Approaches for Efficient Gene Silencing in Plants

Hua Yan, Robert Chretien, Jingsong Ye and Caius M. Rommens*

Simplot Plant Sciences, J.R. Simplot Company, Boise, Idaho 83706

An important component of conventional sense, antisense, and double-strand RNA-based gene silencing constructs is the transcriptional terminator. Here, we show that this regulatory element becomes obsolete when gene fragments are positioned between two oppositely oriented and functionally active promoters. The resulting convergent transcription triggers gene silencing that is at least as effective as unidirectional promoter-to-terminator transcription. In addition to short, variably sized, and nonpolyadenylated RNAs, terminator-free cassette produced rare, longer transcripts that reach into the flanking promoter. These read-through products did not influence the efficacy and expression levels of the neighboring hygromycin phosphotransferase gene. Replacement of gene fragments by promoter-derived sequences further increased the extent of gene silencing. This finding indicates that genomic DNA may be a more efficient target for gene silencing than gene transcripts.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Caius M. Rommens (crommens{at}simplot.com).

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.106.082271.

* Corresponding author; e-mail crommens{at}simplot.com; fax 208–327–3212.

Received April 17, 2006; returned for revision May 26, 2006; accepted June 4, 2006.




This article has been cited by other articles:


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Z. Luo and Z. Chen
Improperly Terminated, Unpolyadenylated mRNA of Sense Transgenes Is Targeted by RDR6-Mediated RNA Silencing in Arabidopsis
PLANT CELL, March 1, 2007; 19(3): 943 - 958.
[Abstract] [Full Text] [PDF]


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