Development of next-generation dengue virus vaccine

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To develop a new DNA vaccine for dengue virus (DV) infection, we have constructed a plasmid-based infectious clone encoding full-length DV-1 cDNA under control of the minimum cytomegalovirus promoter. In our experiments, yields of WT virus 24 hours after plasmid transfection are regularly in the range of 100 to 1000 PFU/ml. Furthermore, DV-1 reporter replicon with luciferase was made by replacing part of the DV-1 structural gene region with the luciferase gene.To improve the reverse genetics for DV, we have developed a homologous recombination-based reverse genetics for DV. Furthermore, we have rescued a chimeric DV, which expresses the prM and E genes from DV type 2 in a DV type 1 genetic background.These improved infectious cDNA clone and reporter replicon systems described here can be employed for studies of the DV replication and pathogenesis and used to develop vaccines, diagnostics, and therapeutics.