Here we investigated the effect of Beauveria bassiana infection on the survival of Aedes aegypti mosquitoes and the modulation of their susceptibility to dengue virus infection. Application of B. bassiana caused a reduction in the life span of A. aegypti and hindered dengue virus replication in the mosquito midgut. Fungus infection induced the expression a variety anti-microbial and dengue virus restriction factor genes. Transient reverse genetic analyses showed that the JAK-STAT pathway is implicated anti-fungal defense of Aedes mosquitoes. Our data suggest that this B. bassiana-mediated anti-dengue activity is likely to be at least partly indirectly mediated through the activation of the mosquito's anti-dengue Toll and JAK-STAT pathways.