Drosophila microRNA dps-2526-3p suppresses dengue virus replication in Aedes aegypti cell
Objective The toll pathway plays an essential role in the inhibition of dengue virus(DENV)replication in Aedes aegypti.Accordingly,the objective of this study was to examine the impact of microRNA on the Toll pathway regulatory genes and its influence on DENV replication in Ae.aegypti.Methods RNAhybrid software prediction and dual-luciferase reporter gene assay were employed to study the binding relationship between dps-miR-2526-3p and Rel1A gene.The real-time fluorescence quantitative PCR(qPCR)was used in the quantification of nuclear and cytoplasmic dps-miR-2526-3p in Wolbachia-carrying(W+)and Wolbachia-free(W-)Ae.aegypti cells via nucleocytoplasmic separation.The miRNA up-regulation assay or miRNA silencing assay combined with virus infection assay were conducted to ascertain the impact of dps-miR-2526-3p on Rel1A gene expression and DENV replication.Results The result of the prediction and dual-luciferase reporter gene assay consistently demonstrated the direct binding relationship between dps-miR-2526-3p and the Rel1A gene.The qPCR result proved the presence of dps-miR-2526-3p in the nucleus and cytoplasm of Ae.aegypti cell.Notably,the expression levels of dps-miR-2526-3p and Rel1A gene in W+cells were significantly higher than those in W-cells.The miRNA function assay indicated that dps-miR-2526-3p could positively regulate Rel1A gene expression.The result of viral infection assay showed that dps-miR-2526-3p exhibited a notable inhibitory effect on DENV replication.Conclusions Ae.aegypti utilizes dps-miR-2526-3p to activate the Toll pathway via Rel1A gene,thereby inhibiting the replication of DENV.The novel molecule with anti-DENV properties was found in this study to offer a promising approach for the development of vector control strategies aimed at prevention and control of DENV transmission.
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