Effect of the noncoding RNA produced by ZIKA virus (ZIKV) on expression of Aedes aegypti genes

We report that ZIKV noncoding RNA (sfRNA) affect expression of mosquito genes upon ZIKV infection. We constructed sfRNA-deficient ZIKV mutant (xrRNA2') and conducted transcriptome-wide gene expression profiling of mosquitos infected with WT and xrRNA2' viruses. We found that sfRNA inhibits apoptosis in the infected tissues of Ae. aegypti by altering expression of mosquito genes that control cell death and survival. Overall design: Aedes aegypti mosquitos were injected intrathoracically with 200 nL of wild type ZIKV Uganda MR766 or xrRNA2' mutant viruses respectively at 5x104 FFU/ml. At 10 days after inoculation mosquitos were euthanised by carbon dioxide exposure, combined into pools of 10 mosquitos in each and subjected to total RNA isolation using TRIreagent (Sigma). Three biological replicates representing three pools of mosquitos were used for library preparation and sequencing on Illumina HiSeq4000 instrument. Libraries were barcoded and 3 of them were loaded into the same lane. To minimize lane-to-lane variations, each library was loaded into 3 different lanes.