Establishment of time- and cell-specific RNAi in Caenorhabditis elegans

Masayuki Hamakawa, Takaaki Hirotsu

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages67-79
Number of pages13
DOIs
Publication statusPublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1507
ISSN (Print)1064-3745

Fingerprint

Caenorhabditis elegans
RNA Interference
Shock
Hot Temperature
Genetic Pleiotropy
Genes
Double-Stranded RNA
Smell
Glutamate Receptors
Motor Neurons
Locomotion
RNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Hamakawa, M., & Hirotsu, T. (2017). Establishment of time- and cell-specific RNAi in Caenorhabditis elegans. In Methods in Molecular Biology (pp. 67-79). (Methods in Molecular Biology; Vol. 1507). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6518-2_6

Establishment of time- and cell-specific RNAi in Caenorhabditis elegans. / Hamakawa, Masayuki; Hirotsu, Takaaki.

Methods in Molecular Biology. Humana Press Inc., 2017. p. 67-79 (Methods in Molecular Biology; Vol. 1507).

Research output: Chapter in Book/Report/Conference proceedingChapter

Hamakawa, M & Hirotsu, T 2017, Establishment of time- and cell-specific RNAi in Caenorhabditis elegans. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1507, Humana Press Inc., pp. 67-79. https://doi.org/10.1007/978-1-4939-6518-2_6
Hamakawa M, Hirotsu T. Establishment of time- and cell-specific RNAi in Caenorhabditis elegans. In Methods in Molecular Biology. Humana Press Inc. 2017. p. 67-79. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6518-2_6
Hamakawa, Masayuki ; Hirotsu, Takaaki. / Establishment of time- and cell-specific RNAi in Caenorhabditis elegans. Methods in Molecular Biology. Humana Press Inc., 2017. pp. 67-79 (Methods in Molecular Biology).
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