RNA-mediated interference (RNAi) is an efficient method for disrupting a specific gene in many organisms, especially C. elegans. Although RNAi is highly specific to one gene, the effect of RNAi could not be limited to one cell or single tissue in the standard RNAi method. It could be very useful for RNAi to be effective in one cell or single tissue for dissecting tissue specific functions of a gene. To establish one cell or single tissue specific RNAi method, we will use the
rde-1, RNAi resistant mutants and its cDNA. It is reported that the
rde-1 gene functions in cell autonomous manner. Taking advantage of this possible cell-autonomous function of
rde-1, we have an idea for one cell or single tissue specific RNAi method. Our hypothesis is that the
rde-1 mutant harboring the
rde-1 cDNA under the control of one cell or tissue specific promoters could be RNAi-sensitive only in one cell or single tissue expressing the
rde-1 cDNA. To examine whether our hypothesis is true or not, we planned construction of the
rde-1 mutant strains expressing the
rde-1 cDNA under the control of a
lin-26 promoter as a hypodermis specific promoter or a
myo-3 promoter as a muscle specific promoter. For investigation of RNAi sensitivity in the single tissue, we will use two types of markers. One is GFP expressing only in the single tissue, and the other is some genes, loss of which causes lethal phenotype in the tissue specific manner. In case of the GFP marker, we will construct the strains expressing NLS::GFP in hypodermis (
lin-26 promoter) or muscle (
myo-3 promoter) and examine whether GFP fluorescence disappears only in the
rde-1 rescued tissue by injection of GFP dsRNA. In addition to the GFP marker, we will confirm our hypothesis by investigating whether RNAi of lethal genes causes lethality only to the
rde-1 mutant strain in which
rde-1 is resucued at the tissue that marker genes function. In this meeting, we will report results of our trials and discuss the possibility of one cell or single tissue specific RNAi method.