Project Number: 5430-32000-003-22
Project Type: Specific Cooperative Agreement

Start Date: Sep 15, 2011
End Date: Sep 14, 2015

Objective:
To identify new molecular targets to inhibit infection and transmission of arboviruses by mosquito vectors of these animal and zoonotic pathogens using RNA interference (RNAi).

Approach:
Utilizing Malaysian MRE16 Sindbis strain that replicates in Aedes aegypti midgut as a model arbovirus to assess molecular inhibition targets, both parties have identified potential but distinct inhibition targets for further investigation. This model system will be further used for initial evaluation of additional novel inhibition targets that have been identified in another recently describe model system. The inhibition approach will be to produce small inhibitory RNA (siRNA) by cloning these genes into a RNA transcription vector and using in vitro transcription to produce double-stranded (ds)RNA. The dsRNA will be then be feed or injected into Aedes aegypti, and the mosquito RNA silencing system will produce siRNA. Mosquitoes will then be challenged by infection with the model virus. The inhibition of infection will be assessed by virus titration, qRT-PCR and immunohistochemistry. The second phase of the proposal will be to evaluate proven targets ability to inhibit Rift Valley fever virus infection of and transmission by mosquitoes once appropriate facilities are available.