title: DNA interference: a simple and efficient gene-silencing system for high-throughput functional analysis in the fern adiantum.
abstract: RNA interference (RNAi) has become a powerful tool for determining gene function and is used in a wide variety of organisms. Since it is necessary to generate double-stranded RNA (dsRNA) as an inducer for RNAi, preparation of RNAi-inducing constructs is somewhat cumbersome and time consuming, especially for the thousands of genes used in a genome-wide analysis. To overcome these problems, we have developed a more convenient gene-silencing method in the fern Adiantum using double-stranded DNA (dsDNA) as a model system for functional analysis in plants. Delivery of dsDNA fragments homologous to an endogenous gene into gametophytic cells can induce sequence-specific gene silencing. As it only requires dsDNA fragments homologous to a target gene, PCR-amplified fragments are enough to trigger gene silencing. Maximum gene silencing efficiencies of >90% have been achieved for transformed plants. In addition, simultaneous transfer of dsDNA fragments corresponding to multiple genes still has a silencing effect for individual genes. We term this approach 'DNA interference'.
authors.affiliation: Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, Hachioji, Tokyo, 192-0397 Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Kawai-Toyooka
authors.forename: Chihiro
authors.initials: C
authors.lastname: Kuramoto
authors.forename: Kyoko
authors.initials: K
authors.lastname: Orui
authors.forename: Kentaro
authors.initials: K
authors.lastname: Motoyama
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Kikuchi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kanegae
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Wada
mesh_terms: D035681:Adiantum / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D015536:Down-Regulation; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D018390:Gene Targeting; D017343:Genes, Plant; D010641:Phenotype; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D015398:Signal Transduction
chemical_list: D010940:Plant Proteins; D004247:DNA