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Open AccessArticle

Development, Characterization, and Application of Two Reporter-Expressing Recombinant Zika Viruses

1
Department of Animal, Dairy, and Veterinary Sciences, College of Agriculture and Applied Sciences, Utah State University, Logan, UT 84322, USA
2
Institute for Antiviral Research, Utah State University, Logan, UT 84322, USA
3
Veterinary Diagnostics and Infectious Diseases, Utah Science Technology and Research, Utah State University, Logan, UT 84341, USA
*
Author to whom correspondence should be addressed.
Viruses 2020, 12(5), 572; https://doi.org/10.3390/v12050572
Received: 3 May 2020 / Revised: 16 May 2020 / Accepted: 18 May 2020 / Published: 22 May 2020
(This article belongs to the Special Issue Flavivirus Replication and Pathogenesis)
Zika virus (ZIKV), a mosquito-borne transplacentally transmissible flavivirus, is an enveloped virus with an ~10.8 kb plus-strand RNA genome that can cause neurological disease. To facilitate the identification of potential antivirals, we developed two reporter-expressing ZIKVs, each capable of expressing an enhanced green fluorescent protein or an improved luminescent NanoLuc luciferase. First, a full-length functional ZIKV cDNA clone was engineered as a bacterial artificial chromosome, with each reporter gene under the cap-independent translational control of a cardiovirus-derived internal ribosome entry site inserted downstream of the single open reading frame of the viral genome. Two reporter-expressing ZIKVs were then generated by transfection of ZIKV-susceptible BHK-21 cells with infectious RNAs derived by in vitro run-off transcription from the respective cDNAs. As compared to the parental virus, the two reporter-expressing ZIKVs grew to lower titers with slower growth kinetics and formed smaller foci; however, they displayed a genome-wide viral protein expression profile identical to that of the parental virus, except for two previously unrecognized larger forms of the C and NS1 proteins. We then used the NanoLuc-expressing ZIKV to assess the in vitro antiviral activity of three inhibitors (T-705, NITD-008, and ribavirin). Altogether, our reporter-expressing ZIKVs represent an excellent molecular tool for the discovery of novel antivirals. View Full-Text
Keywords: Zika virus; infectious cDNA; reporter-expressing virus; green fluorescent protein; luciferase; viral protein expression profile; antiviral Zika virus; infectious cDNA; reporter-expressing virus; green fluorescent protein; luciferase; viral protein expression profile; antiviral
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Yun, S.-I.; Song, B.-H.; Woolley, M.E.; Frank, J.C.; Julander, J.G.; Lee, Y.-M. Development, Characterization, and Application of Two Reporter-Expressing Recombinant Zika Viruses. Viruses 2020, 12, 572.

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