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

SYSNO ASEP	0349491
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	RNA Interference in Schistosoma mansoni Schistosomula: Selectivity, Sensitivity and Operation for Larger-Scale Screening
Author(s)	Štefanič, S. (US) Dvořák, Jan (BC-A) Horn, Martin (UOCHB-X)_{RID, ORCID} Braschi, S. (US) Sojka, Daniel (BC-A)_{RID, ORCID} Ruelas, D. (US) Suzuki, B. (US) Lim, K.-C. (US) Hopkins, S. D. (US) McKerrow, J. H. (US) Caffrey, C. R. (US)
Source Title	PLoS Neglected Tropical Diseases. - : Public Library of Science - ISSN 1935-2735 Roč. 4, č. 10 (2010), e850
Number of pages	16 s.
Language	eng - English
Country	US - United States
Keywords	Tropical disease ; Schistosoma ; schistosomula ; RNAi ; protease ; tegument ; drug target ; drug discovery
Subject RIV	EB - Genetics ; Molecular Biology
R&D Projects	GA203/09/1585 GA ČR - Czech Science Foundation (CSF)
	KJB600960911 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR)
	LC06009 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	ME10011 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
CEZ	AV0Z40550506 - UOCHB-X (2005-2011)
	AV0Z60220518 - PAU-O, BC-A (2005-2011)
UT WOS	000283559600019
DOI	10.1371/journal.pntd.0000850
Annotation	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.
Workplace	Biology Centre (since 2006)
Contact	Dana Hypšová, eje@eje.cz, Tel.: 387 775 214
Year of Publishing	2012

Number of the records: 1