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Published on April 25, 2008; 10.1104/pp.108.118307

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Received February 23, 2008
Accepted April 17, 2008

Artificial trans-acting siRNAs confer consistent and effective gene silencing

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

DuPont Crop Genetics Research, Experimental Station, PO Box 80353, Wilmington, DE 19880-0353, USA

* Corresponding author; email: robert-w2.williams{at}cgr.dupont.com.

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 post-transcriptional gene silencing pathways to block translation of designated transcripts. Here we present a novel gene silencing method utilizing artificial trans-acting small interfering RNAs (ata-siRNAs) in Arabidopsis thaliana. Replacing the endogenous siRNAs 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 a miR167 target sequence led to variable, inefficient silencing of FAD2, suggesting a specific requirement for the miR173 trigger for production of siRNAs from the TAS1c locus.






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