RNA interference targeting cathepsin B of the carcinogenic liver fluke, Opisthorchis viverrini

Publisher:
Elsevier
Publication Type:
Journal Article
Citation:
Parasitology International, 2011, 60 (3), pp. 283 - 288
Issue Date:
2011-01
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Functional genomics have not been reported for Opisthorchis viverrini or the related fish-borne fluke, Clonorchis sinensis.Here we describe the introduction by squarewave electroporation of Cy3-labeled small RNA into adult O. viverrini worms. Adult flukeswere subjected to squarewave electroporation employing a single pulse for 20 msof 125 Vin the presence of 50 ìg/ml of Cy3-siRNA. The parasites tolerated this manipulation and, at 24 and 48 h after electroporation, fluorescence from the Cy3-siRNA was evident throughout the parenchyma of the worms, with strong fluorescence evident in the guts and reproductive organs of the adult worms. Second, other worms were treated using the same electroporation settings with double stranded RNA targeting an endogenous papain-like cysteine protease, cathepsin B. This manipulation resulted in a significant reduction in specific mRNA levels encoding cathepsin B, and a significant reduction in cathepsin B activity against the diagnostic peptide, Z-Arg-Arg-AMC. This appears to be the first report of introduction of reporter genes into O. viverrini and the first report of experimental RNA interference (RNAi) in this fluke. The findings indicated the presence of an intact RNAi pathway in these parasites which, in turn, provides an opportunity to probe gene functions in this neglected tropical disease pathogen.
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