Establishment of Time- and Cell-Specific RNAi in Caenorhabditis elegans.
The nematode worm Caenorhabditis elegans, in which loss-of-function mutants and RNA interference (RNAi) models are available, is a model organism useful for analyzing effects of genes on various life phenomena. In particular, RNAi is a powerful tool that enables time- or cell-specific knockdown via heat shock-inducible RNAi or cell-specific RNAi. However, the conventional RNAi methods are insufficient for investigating pleiotropic genes with various sites of action and life stage-dependent functions. To investigate the temporal- and cell-specific profiles of multifunctional genes, we established a new RNAi method that enables simultaneous time- and cell-specific knockdown (T.C.RNAi) in C. elegans. In this method, one RNA strand is expressed by a cell-specific promoter and the other by a heat shock promoter, resulting in only expression of double-stranded RNA in the target cell when heat shock is induced. We confirmed the effect of T.C.RNAi by the knockdown of GFP and the odr-3 gene which encodes Gα and is essential for olfaction. Further, this technique revealed that the control of glutamate receptors GLR-1 localization in RMD motor neurons requires Ras at the adult stage to regulate locomotion behavior.
Graduate School of Systems Life Sciences, Kyushu University, Higashi-ku, Fukuoka, 812-8581, Japan.
Masayuki
M
Hamakawa
Graduate School of Systems Life Sciences, Kyushu University, Higashi-ku, Fukuoka, 812-8581, Japan. hirotsu.takaaki.056@m.kyushu-u.ac.jp.|Department of Biology, Graduate School of Sciences, Kyushu University, Fukuoka, 812-8581, Japan. hirotsu.takaaki.056@m.kyushu-u.ac.jp.
Takaaki
T
Hirotsu
D000818:Animals; D017173:Caenorhabditis elegans / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D017238:Genes, Helminth; D009928:Organ Specificity; D034622:RNA Interference; D059467:Transcriptome
D029742:Caenorhabditis elegans Proteins
C. elegans; Conditional knockdown; Heat shock; RNAi; Time and cell specific