Počet záznamů: 1

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

  1. 1.
    0349491 - BC-A 2012 RIV US eng J - Článek v odborném periodiku
    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
    Grant CEP: GA ČR GA203/09/1585; GA AV ČR KJB600960911; GA MŠk(CZ) LC06009; GA MŠk(CZ) ME10011
    Výzkumný záměr: CEZ:AV0Z40550506; CEZ:AV0Z60220518
    Klíčová slova: Tropical disease * Schistosoma * schistosomula * RNAi * protease * tegument * drug target * drug discovery
    Kód oboru RIV: EB - Genetika a molekulární biologie
    Impakt faktor: 4.752, rok: 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.
    Trvalý link: http://hdl.handle.net/11104/0189712