Silencing CRVP379 inhibits DENV acquisition in live mosquitoes.

A-E. Mosquitoes were intra-thoracically injected with either dsRNA against the coding region of CRVP379 or dsRNA against GFP as control. At 4 dpmi, mosquitoes were infected with DENV through blood feeding. At 4 dpi, midgut tissues were dissected and individually analyzed for gene expression with qRT-PCR analysis. Each data point represents one mosquito midgut. A. Levels of CRVP379 in select midguts where RNAi was successful, as compared to levels in control mosquitoes. P<0.01. B. Levels of DENV infection in select midguts where RNAi was successful, as compared to levels in control mosquitoes. P<0.01. Infection rates in midguts where CRVP379 RNAi was successful ranged from to .000000765–.0000315 ng DENV E/ng actin. C. Levels of CRVP379 in midguts where RNAi was both successful and unsuccessful, as compared to levels in control mosquitoes. D. Levels of DENV infection in midguts where RNAi was both successful and unsuccessful, as compared to levels in control mosquitoes. C-D. Squares represent midguts where RNAi did not knock down CRVP379 successfully, cirlces represent midguts where RNAi did knock down CRVP379 successfully. E. Levels of DENV infection correspond to levels of CRVP379 expression. Both midguts where RNAi did and did not knock down CRVP379 were analyzed for both CRVP379 expression and DENV infection by qRT-PCR. Data is plotted as ngs of DENV E versus levels of CRVP379, normalized to mosquito actin. Data correlated with Pearson, r = 0.6442, P<0.0001 F. Silencing CRVP379 reduces DENV infection in whole mosquitoes. Mosquitoes were intra-thoracically injected with either dsRNA against the coding region of CRVP379 or dsRNA against GFP as control. At 4 dpmi, mosquitoes were infected with DENV through blood feeding. At 7 dpi, homogenized whole mosquitoes were individually analyzed for gene expression with qRT-PCR analysis.