[Construction of AalDV-3 Mediated mirRNA Sponge Delivery System and Its Effect on Cell and Larva of Aedes aegypti].

Abstract

microRNAs (miRNAs) not only play the key roles in regulation of the growth and development of mosquito, but also has an important function in interaction between the pathogen and vector. So, miRNAs can be used as the molecular target for development of an alternative method for mosquito and mosquito-borne disease control. However, an effective delivery system is still need. Mosquito densovirus have the potential for vector control as transducing agents to express foreign toxins or small interfering RNAs molecules in vitro and in vivo. In this study, we report the development of a recombinant Aedes albopictus densovirus-3(AalDV-3) miRNA sponge expression system, using an intronic miRNA sponge expression strategy. To test the inhibition effect of recombinant virus medicated miRNA sponge on endogenous miR-210, Aedes aegypti Aag2 cell and 1st-2nd larvae were infected, respectively. The splicing of the intronic miRNA sponge expression constructs in vitro and in vivo were tested by RT-PCR with intron span primers, and the relatively expression level of miR-210 was confirmed by qPCR. As results,AalDV-3can be used to decrease endogenous miRNAs by generating an antisense sponge in vitro and in vivo, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of densovirus for vector control.