title: Simple gene silencing using the trans-acting siRNA pathway.
abstract: In plants, particular micro-RNAs (miRNAs) induce the production of a class of small interfering RNAs (siRNA) called trans-acting siRNA (ta-siRNA) that lead to gene silencing. A single miRNA target is sufficient for the production of ta-siRNAs, which target can be incorporated into a vector to induce the production of siRNAs, and ultimately gene silencing. The term miRNA-induced gene silencing (MIGS) has been used to describe such vector systems in Arabidopsis. Several ta-siRNA loci have been identified in soybean, but, prior to this work, few of the inducing miRNAs have been experimentally validated, much less used to silence genes. Nine ta-siRNA loci and their respective miRNA targets were identified, and the abundance of the inducing miRNAs varies dramatically in different tissues. The miRNA targets were experimentally verified by silencing a transgenic GFP gene and two endogenous genes in hairy roots and transgenic plants. Small RNAs were produced in patterns consistent with the utilization of the ta-siRNA pathway. A side-by-side experiment demonstrated that MIGS is as effective at inducing gene silencing as traditional hairpin vectors in soybean hairy roots. Soybean plants transformed with MIGS vectors produced siRNAs and silencing was observed in the T1 generation. These results complement previous reports in Arabidopsis by demonstrating that MIGS is an efficient way to produce siRNAs and induce gene silencing in other species, as shown with soybean. The miRNA targets identified here are simple to incorporate into silencing vectors and offer an effective and efficient alternative to other gene silencing strategies.
authors: Institute for Plant Breeding, Genetics and Genomics, University of Georgia, Athens, GA, USA.|Center for Applied Genetic Technologies, University of Georgia, Athens, GA, USA.
authors: Thomas B
authors: TB
authors: Jacobs
authors: Center for Applied Genetic Technologies, University of Georgia, Athens, GA, USA.
authors: Noah J
authors: NJ
authors: Lawler
authors: Center for Applied Genetic Technologies, University of Georgia, Athens, GA, USA.|Department of Crop and Soil Sciences, University of Georgia, Athens, GA, USA.
authors: Peter R
authors: PR
authors: LaFayette
authors: Department of Crop Sciences, University of Illinois, Urbana, IL, USA.
authors: Lila O
authors: LO
authors: Vodkin
authors: Institute for Plant Breeding, Genetics and Genomics, University of Georgia, Athens, GA, USA.|Center for Applied Genetic Technologies, University of Georgia, Athens, GA, USA.|Department of Crop and Soil Sciences, University of Georgia, Athens, GA, USA.
authors: Wayne A
authors: WA
authors: Parrott
mesh_terms: D001483:Base Sequence; D020868:Gene Silencing; D017343:Genes, Plant; D005822:Genetic Vectors / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics; D030821:Plants, Genetically Modified; D034741:RNA, Small Interfering / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013025:Glycine max / Q000235:genetics
chemical_list: D035683:MicroRNAs; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins
keywords: Plant biotechnology; functional genomics; gene silencing; transgenic crops