Development of a replicon cell line-based high throughput antiviral assay for screening inhibitors of Zika virus

Antiviral Research
Jia-Qi LiHanqing Ye

Abstract

Zika virus (ZIKV) is an important emerging human pathogen associated with microcephaly, Guillain-Barré syndrome and meningoencephalitis. Developing rapid and reliable HTS assay is important for ZIKV drug discovery. Here, we constructed a dicistronic ZIKV replicon (ZIKV-Pac-Rluc-Rep) that contained the Renilla luciferase (Rluc) reporter gene separated from the puromycin N-acetyl-transferase (Pac) selectable marker by a short peptide cleavage site. A clonal replicon cell line stably expressing high level of ZIKV replicon was established by selection with puromycin. By optimizing cell number, compound concentration and incubation time, a robust replicon cell-based HTS assay was developed with a calculated Z' value of >0.5. The fully optimized assay was further validated using several known flavivirus replication inhibitors. Altogether, the replicon cell-based HTS assay developed in this study will facilitate the discovery of antiviral compounds against ZIKV.

Citations

Oct 17, 2018·The Journal of General Virology·Lindomar José PenaAntonio Carlos de Freitas
Dec 24, 2019·The Journal of Cell Biology·Yali CiLei Shi
Dec 24, 2018·Viruses·Chaker El KalamouniWildriss Viranaïcken
Jul 31, 2019·Nucleic Acids Research·Shan XuLei Shi
Nov 6, 2018·Viruses·Ginés Ávila-PérezLuis Martínez-Sobrido
Jun 22, 2020·Current Opinion in Virology·Camila R Fontes-GarfiasPei-Yong Shi
May 14, 2021·Journal of Virology·Xiaoying JiaWei Wang

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