Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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First published online April 25, 2008; 10.1104/pp.108.118307

Plant Physiology 147:543-551 (2008)
© 2008 American Society of Plant Biologists

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BREAKTHROUGH TECHNOLOGIES

Artificial trans-Acting siRNAs Confer Consistent and Effective Gene Silencing

Maria de la Luz Gutiérrez-Nava, Milo J. Aukerman, Hajime Sakai, Scott V. Tingey and Robert W. Williams*

DuPont Crop Genetics Research, Experimental Station, Wilmington, Delaware 19880–0353

Manipulating gene expression is critical to exploring gene function and a useful tool for altering commercial traits. Techniques such as hairpin-based RNA interference, virus-induced gene silencing, and artificial microRNAs take advantage of endogenous posttranscriptional gene silencing pathways to block translation of designated transcripts. Here we present a novel gene silencing method utilizing artificial trans-acting small interfering RNAs in Arabidopsis (Arabidopsis thaliana). Replacing the endogenous small interfering RNAs encoded in the TAS1c gene with sequences from the FAD2 gene silenced FAD2 activity to levels comparable to the fad2-1 null allele in nearly all transgenic events. Interestingly, exchanging the endogenous miR173 target sequence in TAS1c with an miR167 target sequence led to variable, inefficient silencing of FAD2, suggesting a specific requirement for the miR173 trigger for production of small interfering RNAs from the TAS1c locus.

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: Robert W. Williams (robert-w2.williams{at}cgr.dupont.com).

www.plantphysiol.org/cgi/doi/10.1104/pp.108.118307

* Corresponding author; e-mail robert-w2.williams{at}cgr.dupont.com.

Received February 23, 2008; accepted April 17, 2008; published April 25, 2008.

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