title: RNA-mediated epigenetic modifications of an endogenous gene targeted by a viral vector: a potent gene silencing system to produce a plant that does not carry a transgene but has altered traits.
abstract: Nucleotide-sequence-specific interactions mediated by double-stranded RNA (dsRNA) can induce gene silencing. Gene silencing through transcriptional repression can be induced by dsRNA targeted to a gene promoter. However, until recently, no plant has been produced that harbors an endogenous gene that remains silenced in the absence of promoter-targeting dsRNA. We have reported for the first time that transcriptional gene silencing can be induced by targeting dsRNA to the endogenous gene promoters in petunia and tomato plants, using a Cucumber mosaic virus (CMV)-based vector and that the induced gene silencing is heritable. Efficient silencing depended on the function of the 2b protein encoded in the vector, which facilitates epigenetic modifications through the transport of short interfering RNA (siRNA) to the nucleus. Here we show that gene silencing that is mediated by targeting dsRNA to a gene promoter via the CMV vector can be as strong as co-suppression in terms of both the extent of mRNA decrease and phenotypic changes. We also show that the expression of genes involved in RNA-directed DNA methylation and in demethylation are upregulated and downregulated, respectively, in Arabidopsis plants infected with CMV. Thus, along with the function of the 2b protein, that transports siRNA to the nucleus, the promoter-targeted silencing system using the CMV vector has some property that facilitates heritable epigenetic changes on endogenous genes, enabling the production of a novel class of modified plants that do not have a transgene.
authors.affiliation: Research Faculty of Agriculture, Hokkaido University, Sapporo, Japan. kanazawa@res.agr.hokudai.ac.jp
authors.forename: Akira
authors.initials: A
authors.lastname: Kanazawa
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Inaba
authors.forename: Megumi
authors.initials: M
authors.lastname: Kasai
authors.forename: Hanako
authors.initials: H
authors.lastname: Shimura
authors.forename: Chikara
authors.initials: C
authors.lastname: Masuta
mesh_terms: D017799:Cucumovirus / Q000235:genetics; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D005822:Genetic Vectors; D032306:Petunia / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D011401:Promoter Regions, Genetic; D012330:RNA, Double-Stranded / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics*; D019076:Transgenes; D014764:Viral Proteins / Q000235:genetics
chemical_list: C404227:2b protein, cucumber mosaic virus; D012330:RNA, Double-Stranded; D014764:Viral Proteins