Characterization of Recombinant Flaviviridae Viruses Possessing a Small Reporter Tag

Journal of Virology
Tomokazu TamuraYoshiharu Matsuura

Abstract

The family Flaviviridae consists of four genera, Flavivirus, Pestivirus, Pegivirus, and Hepacivirus, and comprises important pathogens of human and animals. Although the construction of recombinant viruses carrying reporter genes encoding fluorescent and bioluminescent proteins has been reported, the stable insertion of foreign genes into viral genomes retaining infectivity remains difficult. Here, we applied the 11-amino-acid subunit derived from NanoLuc luciferase to the engineering of the Flaviviridae viruses and then examined the biological characteristics of the viruses. We successfully generated recombinant viruses carrying the split-luciferase gene, including dengue virus, Japanese encephalitis virus, hepatitis C virus (HCV), and bovine viral diarrhea virus. The stability of the viruses was confirmed by five rounds of serial passages in the respective susceptible cell lines. The propagation of the recombinant luciferase viruses in each cell line was comparable to that of the parental viruses. By using a purified counterpart luciferase protein, this split-luciferase assay can be applicable in various cell lines, even when it is difficult to transduce the counterpart gene. The efficacy of antiviral reagents against the rec...Continue Reading

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Citations

Mar 3, 2018·Viruses·Michael J McFaddenStacy M Horner
Jul 20, 2019·PLoS Computational Biology·James J BullRustom Antia
Aug 4, 2019·Journal of Virology·Yutaka TeradaWataru Kamitani
Aug 30, 2019·Journal of Virology·Tomokazu TamuraYoshiharu Matsuura
Aug 23, 2020·International Journal of Molecular Sciences·Natasa GasparLaura Mezzanotte
Oct 1, 2020·Viruses·Coleman Baker, Pei-Yong Shi
Apr 14, 2019·Scientific Reports·Christiane RiedelTill Rümenapf
Sep 28, 2020·Antiviral Research·Ricardo Sanchez-VelazquezMargus Varjak
Jan 14, 2021·Virus Research·Yuta Kanai, Takeshi Kobayashi
Jun 26, 2021·Current Opinion in Virology·Carlos Daniel Cordero-RiveraRosa María Del Ángel
Aug 11, 2020·Virology·Hironobu MurakamiKenji Tsukamoto
Jul 10, 2021·Virology·Hironobu MurakamiKenji Tsukamoto

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