title: Artificial trans-acting siRNAs confer consistent and effective gene silencing.
abstract: 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.
authors.affiliation: DuPont Crop Genetics Research, Experimental Station, Wilmington, Delaware 19880-0353, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: de la Luz Gutiérrez-Nava
authors.forename: Milo J
authors.initials: MJ
authors.lastname: Aukerman
authors.forename: Hajime
authors.initials: H
authors.lastname: Sakai
authors.forename: Scott V
authors.initials: SV
authors.lastname: Tingey
authors.forename: Robert W
authors.initials: RW
authors.lastname: Williams
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers; D044943:Fatty Acid Desaturases / Q000235:genetics; D020868:Gene Silencing; D017343:Genes, Plant; D030821:Plants, Genetically Modified; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D017931:DNA Primers; D034741:RNA, Small Interfering; D044943:Fatty Acid Desaturases; C523310:FAD2 protein, Arabidopsis