Gene silencing in the spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis>: dsRNA and siRNA parental silencing of the <Emphasis Type="Italic">Distal-less</Emphasis> gene |
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Authors: | Abderrahman Khila Miodrag Grbi? |
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Institution: | (1) Department of Biology, University of Western Ontario, London, N6A 5B7, Canada;(2) Department of Biology, McGill University, Montreal, Quebec, H3A 1B1, Canada |
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Abstract: | A major prerequisite to understanding the evolution of developmental programs includes an appreciation of gene function in
a comparative context. RNA interference (RNAi) represents a powerful method for reverse genetics analysis of gene function.
However, RNAi protocols exist for only a handful of arthropod species. To extend functional analysis in basal arthropods,
we developed a RNAi protocol for the two-spotted spider mite Tetranychus urticae focusing on Distal-less (Dll), a conserved gene involved in appendage specification in metazoans. First, we describe limb morphogenesis in T. urticae using confocal and scanning electron microscopy. Second, we examine T. urticae Dll (Tu-Dll) mRNA expression patterns and correlate its expression with appendage development. We then show that fluorescently labeled
double-stranded RNA (dsRNA) and short interfering RNA (siRNA) molecules injected into the abdomen of adult females are incorporated
into the oviposited eggs, suggesting that dsRNA reagents can be systemically distributed in spider mites. Injection of longer
dsRNA as well as siRNA induced canonical limb truncation phenotypes as well as the fusion of leg segments. Our data suggest
that Dll plays a conserved role in appendage formation in arthropods and that such conserved genes can serve as reliable starting
points for the development of functional protocols in nonmodel organisms. |
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Keywords: | Chelicerata Embryo development RNAi Evolution of development Limb development Distal-less |
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