Establishment of a tissue-specific RNAi system in C. elegans.
In C. elegans, mosaic analysis is a powerful genetic tool for determining in which tissue or specific cells a gene of interest is required. For traditional mosaic analysis, a loss-of-function mutant and a genomic fragment that can rescue the mutant phenotype are required. Here we establish an easy and rapid mosaic system using RNAi (RNA mediated interference), using a rde-1 mutant that is resistant to RNAi. Tissue-specific expression of the wild type rde-1 cDNA in rde-1 mutants limits RNAi sensitivity to a specific tissue. We established hypodermal-and muscle-specific RNAi systems by expressing rde-1 cDNA under the control of the lin-26 and hlh-1 promoters, respectively. We confirmed tissue-specific RNAi using two assays: (1) tissue-specific knockdown of GFP expression, and (2) phenocopy of mutations in essential genes that were previously known to function in a tissue-specific manner. We also applied this system to an essential gene, ajm-1, expressed in hypodermis and gut, and show that lethality in ajm-1 mutants is due to loss of expression in hypodermal cells. Although we demonstrate tissue-specific RNAi in hypodermis and muscle, this method could be easily applied to other tissues.
Qadota
Hiroshi
H
Division of Signal Transduction, Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Nara 630-0101, Japan. hkadota@emory.edu
Inoue
Makiko
M
Hikita
Takao
T
Köppen
Mathias
M
Hardin
Jeffrey D
JD
Amano
Mutsuki
M
Moerman
Donald G
DG
Kaibuchi
Kozo
K
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics* / Q000502:physiology; D005804:Genes, Lethal; D009132:Muscles / Q000378:metabolism; D009154:Mutation; D009928:Organ Specificity; D010641:Phenotype; D011401:Promoter Regions, Genetic; D034622:RNA Interference
C441580:AJM-1 protein, C elegans; D029742:Caenorhabditis elegans Proteins; C122185:rde-1 protein, C elegans