Effective inhibition of different Japanese encephalitis virus genotypes by RNA interference targeting two conserved viral gene sequences in vitro and in vivo
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Japanese encephalitis is a zoonotic, mosquito-borne, infectious disease caused by Japanese encephalitis virus (JEV), which is prevalent in China. At present, there are no specific drugs or therapies for JEV infection, which can only be treated symptomatically. Lentivirus-mediated RNA interference (RNAi) is a highly efficient method to silence target genes. In this study, two lentiviral shRNA, LV-C and LV-NS5, targeting the conserved viral gene sequences were used to inhibit different JEV genotypes strains in BHK21 cells and mice. The results showed that LV-C significantly inhibited JEV genotype I and genotype III strains in cells and mice. Quantitative RT-PCR analysis showed that JEV mRNA were reduced by 83.2–90.9% in cells by LV-C and that flow cytometry analysis confirmed the inhibitory activity of LV-C. The viral titers were reduced by about 1000-fold in cells and the brains of suckling mice by LV-C, and the pretreatment of LV-C protected 60–80% of mice against JEV-induced lethality. The inhibitory activities of LV-NS5 in cells and mice were weaker than those of LV-C. These results indicate that RNAi targeting of the two conserved viral gene sequences had significantly suppressed the replication of different JEV genotypes strains in vitro and in vivo, highlighting the feasibility of RNAi targeting of conserved viral gene sequences for controlling JEV infection.
KeywordsJapanese encephalitis virus Conserved sequence Genotype Lentiviral shRNA Antiviral
This work was supported by the Educational Commission of SIchuan Province of China (No. 18ZA0206) and the Research Fund for the Doctoral Program of North Sichuan Medical College (No. CBY16-QD03).
LY, XJF, and JY conceived and designed the study. CYL, PL, GLY, and HR collected the samples. LY, XLG, and YL carried out the experiments. XJF, RH, and YLF performed the data analysis. XJF wrote the first draft. LY and JY reviewed and edited the manuscript. All authors read and approved the final manuscript.
Compliance with ethical standards
Conflict of interest
All authors declared that they have no conflicts of interest to this work.
All procedures performed in this study involving animals were approved by the Institutional Animal Care and Use Committee of North Sichuan Medical College (Approval Number BM2017-056), Nanchong city, China and followed National Institutes of Health guidelines.
Informed consent was obtained from all individual participants included in the study.