Journal Article

DNA Interference: a Simple and Efficient Gene-Silencing System for High-Throughput Functional Analysis in the Fern <i>Adiantum</i>

Hiroko Kawai-Toyooka, Chihiro Kuramoto, Kyoko Orui, Kentaro Motoyama, Kazuhiro Kikuchi, Takeshi Kanegae and Masamitsu Wada

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 45, issue 11, pages 1648-1657
Published in print November 2004 | ISSN: 0032-0781
Published online November 2004 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pch186
DNA Interference: a Simple and Efficient Gene-Silencing System for High-Throughput Functional Analysis in the Fern Adiantum

More Like This

Show all results sharing these subjects:

  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

GO

Show Summary Details

Preview

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’.

Keywords: Keywords: Adiantum capillus-veneris — DNAi — dsDNA — Fern — Gene silencing — RNAi.; Abbreviations: CaMV, cauliflower mosaic virus; DNAi, DNA interference; dsDNA, double-stranded DNA; dsRNA, double-stranded RNA; EST, expressed sequence tag; GFP, green fluorescent protein; hpRNA, hairpin RNA; RNAi, RNA interference; RT-PCR, reverse transcription-PCR.

Journal Article.  6043 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.