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

Sripa, J; Pinlaor, P; Brindley, PJ; Sripa, B; Kaewkes, S; Robinson, MW; Young, ND; Gasser, RB; Loukas, A; Laha, T

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

Sripa, J Pinlaor, P Brindley, PJ Sripa, B Kaewkes, S Robinson, MW

Young, ND Gasser, RB Loukas, A Laha, T

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Publication Type:: Journal Article
Citation:: Parasitology International, 2011, 60 (3), pp. 283 - 288
Issue Date:: 2011-09-01

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Field	Value	Language
dc.contributor.author	Sripa, J	en_US
dc.contributor.author	Pinlaor, P	en_US
dc.contributor.author	Brindley, PJ	en_US
dc.contributor.author	Sripa, B	en_US
dc.contributor.author	Kaewkes, S	en_US
dc.contributor.author	Robinson, MW https://orcid.org/0000-0001-7191-0034	en_US
dc.contributor.author	Young, ND	en_US
dc.contributor.author	Gasser, RB	en_US
dc.contributor.author	Loukas, A	en_US
dc.contributor.author	Laha, T	en_US
dc.date.available	2011-04-21	en_US
dc.date.issued	2011-09-01	en_US
dc.identifier.citation	Parasitology International, 2011, 60 (3), pp. 283 - 288	en_US
dc.identifier.issn	1383-5769	en_US
dc.identifier.uri	http://hdl.handle.net/10453/18126
dc.description.abstract	Functional genomics have not been reported for Opisthorchis viverrini or the related fish-borne fluke, Clonorchis sinensis. Here we describe the introduction by square wave electroporation of Cy3-labeled small RNA into adult O. viverrini worms. Adult flukes were subjected to square wave electroporation employing a single pulse for 20. ms of 125. V in the presence of 50. g/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. © 2011 Elsevier Ireland Ltd.	en_US
dc.relation.ispartof	Parasitology International	en_US
dc.relation.isbasedon	10.1016/j.parint.2011.04.003	en_US
dc.subject.classification	Mycology & Parasitology	en_US
dc.subject.mesh	Bile Ducts, Intrahepatic	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Mesocricetus	en_US
dc.subject.mesh	Opisthorchis	en_US
dc.subject.mesh	Opisthorchiasis	en_US
dc.subject.mesh	Cholangiocarcinoma	en_US
dc.subject.mesh	Bile Duct Neoplasms	en_US
dc.subject.mesh	Cathepsin B	en_US
dc.subject.mesh	Intercellular Signaling Peptides and Proteins	en_US
dc.subject.mesh	Helminth Proteins	en_US
dc.subject.mesh	RNA, Double-Stranded	en_US
dc.subject.mesh	RNA, Helminth	en_US
dc.subject.mesh	RNA, Messenger	en_US
dc.subject.mesh	Electroporation	en_US
dc.subject.mesh	Transduction, Genetic	en_US
dc.subject.mesh	RNA Interference	en_US
dc.subject.mesh	Time Factors	en_US
dc.subject.mesh	Cricetinae	en_US
dc.subject.mesh	Progranulins	en_US
dc.title	RNA interference targeting cathepsin B of the carcinogenic liver fluke, Opisthorchis viverrini	en_US
dc.type	Journal Article
utslib.citation.volume	3	en_US
utslib.citation.volume	60	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	0707 Veterinary Sciences	en_US
utslib.for	1108 Medical Microbiology	en_US
dc.location.activity	Netherlands
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
utslib.copyright.status	closed_access
pubs.issue	3	en_US
pubs.publication-status	Published	en_US
pubs.volume	60	en_US

Abstract:

Functional genomics have not been reported for Opisthorchis viverrini or the related fish-borne fluke, Clonorchis sinensis. Here we describe the introduction by square wave electroporation of Cy3-labeled small RNA into adult O. viverrini worms. Adult flukes were subjected to square wave electroporation employing a single pulse for 20. ms of 125. V in the presence of 50. g/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. © 2011 Elsevier Ireland Ltd.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/18126