BBSRC-Pfizer student: Analysis of an anti-silencing mechanism involved in immune evasion by vector borne dsRNA animal viruses of family Reoviridae

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The reoviruses have 9 to 12 segmented dsRNA genomes and include important animal pathogens (including humans). The project will investigate the mechanisms these viruses use to evade RNA silencing. dsRNA-binding/helicase proteins from Bluetongue and Banna viruses, will be expressed to investigate their ability to inactivate silencing by sequestering/unwinding 21bp dsRNA silencing molecules. Antibodies to these proteins (e.g. Banna virus VP12 and orthoreovirus sigma 3) will be used to identify their sub-cellular location (confocal microscopy). Gel shift assays (using siRNAs) will be used to confirm dsRNA binding by the expressed proteins. Expression of native proteins in the cell will itself be subjected to silencing, by pre-treating cells with appropriate siRNAs. The same cells will then be transfected (TAT-mediated transfection) with anti-silencing proteins from other viruses (e.g. tombusvirus p19 protein), to elucidate the antisilencing mechanism involved

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