@article{mbs:/content/journal/jgv/10.1099/vir.0.058008-0, author = "Suzuki, Ryosuke and Ishikawa, Tomohiro and Konishi, Eiji and Matsuda, Mami and Watashi, Koichi and Aizaki, Hideki and Takasaki, Tomohiko and Wakita, Takaji", title = "Production of single-round infectious chimeric flaviviruses with DNA-based Japanese encephalitis virus replicon", journal= "Journal of General Virology", year = "2014", volume = "95", number = "1", pages = "60-65", doi = "https://doi.org/10.1099/vir.0.058008-0", url = "https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.058008-0", publisher = "Microbiology Society", issn = "1465-2099", type = "Journal Article", abstract = "A method for rapid production of single-round infectious particles (SRIPs) of flavivirus would be useful for viral mutagenesis studies. Here, we established a DNA-based production system for SRIPs of flavivirus. We constructed a Japanese encephalitis virus (JEV) subgenomic replicon plasmid, which lacked the C-prM-E (capsid–pre-membrane–envelope) coding region, under the control of the cytomegalovirus promoter. When the JEV replicon plasmid was transiently co-transfected with a JEV C-prM-E expression plasmid into 293T cells, SRIPs were produced, indicating successful trans-complementation with JEV structural proteins. Equivalent production levels were observed when C and prM–E proteins were provided separately. Furthermore, dengue types 1–4, West Nile, yellow fever or tick-borne encephalitis virus prM-E proteins could be utilized for production of chimaeric flavivirus SRIPs, although the production was less efficient for dengue and yellow fever viruses. These results indicated that our plasmid-based system is suitable for investigating the life cycles of flaviviruses, diagnostic applications and development of safer vaccine candidates.", }