RNA Interference in Schistosoma mansoni Schistosomula: Selectivity, Sensitivity and Operation for Larger-Scale Screening

0349491 - BC 2012 RIV US eng J - Journal Article
Štefanič, S. - Dvořák, Jan - Horn, Martin - Braschi, S. - Sojka, Daniel - Ruelas, D. - Suzuki, B. - Lim, K.-C. - Hopkins, S. D. - McKerrow, J. H. - Caffrey, C. R.
RNA Interference in Schistosoma mansoni Schistosomula: Selectivity, Sensitivity and Operation for Larger-Scale Screening.
PLoS Neglected Tropical Diseases. Roč. 4, č. 10 (2010), e850. ISSN 1935-2735. E-ISSN 1935-2735
R&D Projects: GA ČR GA203/09/1585; GA AV ČR KJB600960911; GA MŠMT(CZ) LC06009; GA MŠMT(CZ) ME10011
Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z60220518
Keywords : Tropical disease * Schistosoma * schistosomula * RNAi * protease * tegument * drug target * drug discovery
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 4.752, year: 2010

RNA interference (RNAi) is a technique to selectively suppress mRNA of individual genes and, consequently, their cognate proteins. In consideration of large-scale screens to identify candidate drug targets, we examined the selectivity and sensitivity (the degree of suppression) of RNAi for 11 genes produced in different tissues of the parasite: the gut, tegument (surface) and otherwise. We used the schistosomulum stage prepared from infective cercariae larvae which are accessible in large numbers and adaptable to automated screening platforms. We found that RNAi suppresses transcripts selectively, however, the sensitivity of suppression varies. Additionally, we defined operational parameters to facilitate large-scale RNAi, including choice of culture medium, transfection strategy to deliver dsRNA, dose- and time-dependency, and dosing limits. Finally, using fluorescent probes, we show that the developing gut allows rapid entrance of dsRNA into the parasite to initiate RNAi.
Permanent Link: http://hdl.handle.net/11104/0189712