Virus Genes

, Volume 54, Issue 6, pp 746–755 | Cite as

Effective inhibition of different Japanese encephalitis virus genotypes by RNA interference targeting two conserved viral gene sequences in vitro and in vivo

  • Lei Yuan
  • Xiaojuan Feng
  • Xuelian Gao
  • Yu Luo
  • Chaoyue Liu
  • Peng Liu
  • Guolin Yang
  • Hong Ren
  • Rong Huang
  • Yalan Feng
  • Jian YangEmail author


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.


Japanese 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).

Author contributions

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.

Ethics statement

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

Informed consent was obtained from all individual participants included in the study.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Lei Yuan
    • 1
  • Xiaojuan Feng
    • 2
  • Xuelian Gao
    • 3
  • Yu Luo
    • 3
  • Chaoyue Liu
    • 1
  • Peng Liu
    • 1
  • Guolin Yang
    • 4
  • Hong Ren
    • 4
  • Rong Huang
    • 1
  • Yalan Feng
    • 1
  • Jian Yang
    • 1
    Email author
  1. 1.Pathogen and Immunology Experiment Teaching CenterNorth Sichuan Medical CollegeNanchongChina
  2. 2.Medical Functional Experiment Teaching CenterNorth Sichuan Medical CollegeNanchongChina
  3. 3.Department of Medical ImagingNorth Sichuan Medical CollegeNanchongChina
  4. 4.Laboratory Animal CenterNorth Sichuan Medical CollegeNanchongChina

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