Abstract
Japanese encephalitis virus (JEV), an important pathogen in Eastern and Southern Asia and the Pacific, has spread to Australia and other territories in recent years. Although the vaccine for JEV has been used in some countries, development of efficient antiviral drugs is still an urgent requirement. Replicon systems have been widely used in the research of viral replication and antiviral screening for West Nile virus (WNV), yellow fever virus (YFV) and dengue virus (DENV). Here, a novel JEV replicon harboring the Rluc and Pac gene (JEV-Pac-Rluc-Rep) was constructed. Furthermore, we established a BHK-21 cell line harboring JEV-Pac-Rluc-Rep (BHK-21/PAC/Rluc cell line) through continuous puromycin selection. Characterization of cell line stability showed that the replicon RNA could persistently replicate in this cell line for at least up to 10 rounds of passage. Using a known flavivirus inhibitor, the JEV replicon cell line was validated for antiviral screening. The JEV replicon cell line will be a valuable tool for both compound screening and viral replication studies.
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Acknowledgements
We thank the Center for Instrumental Analysis and Metrology, Wuhan Institute of Virology and Wuhan Key Laboratory on Emerging Infectious Diseases and Biosafety for helpful assistance during the course of the work.
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This study was funded by the National Key Research and Development Program of China (2016YFD0500400 to B.Z. and 2016YFC1200400 to H.Q.Y.), the National Natural Science Foundation of China (81572003 to B.Z.).
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The authors declare no conflicts of interest.
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This article does not contain any studies with human participants or animals performed by any of the authors.
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Zhang, QY., Li, XD., Liu, SQ. et al. Development of a stable Japanese encephalitis virus replicon cell line for antiviral screening. Arch Virol 162, 3417–3423 (2017). https://doi.org/10.1007/s00705-017-3508-9
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DOI: https://doi.org/10.1007/s00705-017-3508-9