期刊
VIRUSES-BASEL
卷 12, 期 5, 页码 -出版社
MDPI
DOI: 10.3390/v12050572
关键词
Zika virus; infectious cDNA; reporter-expressing virus; green fluorescent protein; luciferase; viral protein expression profile; antiviral
类别
资金
- Utah Science Technology and Research [A34637, A36815]
- American Board of Obstetrics and Gynecology
- American College of Obstetricians and Gynecologists
- American Society for Reproductive Medicine
- Society for Reproductive Endocrinology and Infertility [200784-00001]
- Utah Agricultural Experiment Station [UTAO-1345]
Zika virus (ZIKV), a mosquito-borne transplacentally transmissible flavivirus, is an enveloped virus with an similar to 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.
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