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.
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
Hiroshi
H
Qadota
Makiko
M
Inoue
Takao
T
Hikita
Mathias
M
Köppen
Jeffrey D
JD
Hardin
Mutsuki
M
Amano
Donald G
DG
Moerman
Kozo
K
Kaibuchi
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