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Published on June 9, 2006; 10.1104/pp.106.082271

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Received April 17, 2006
Returned for revision May 1, 2006
Accepted June 4, 2006

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 ID 83706

* Corresponding author; email: crommens{at}simplot.com.

An important component of conventional sense, antisense, and double-strand (ds) 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 non-polyadenylated 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 (hptII) 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.




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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.
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