Format

Send to

Choose Destination
Gene. 2016 Dec 15;594(2):183-189. doi: 10.1016/j.gene.2016.08.055. Epub 2016 Sep 1.

The NS3 and NS4A genes as the targets of RNA interference inhibit replication of Japanese encephalitis virus in vitro and in vivo.

Author information

1
Research Center of Swine Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China; Laboratory of Zoonosis, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.
2
Research Center of Swine Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China; Laboratory of Zoonosis, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China; Sichuan Science-observation Experiment of Veterinary Drugs and Veterinary Biological Technology, Ministry of Agriculture, Chengdu 611130, China.
3
Research Center of Swine Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China; Laboratory of Zoonosis, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China; Sichuan Science-observation Experiment of Veterinary Drugs and Veterinary Biological Technology, Ministry of Agriculture, Chengdu 611130, China. Electronic address: veterinary226@163.com.

Abstract

Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that can cause acute encephalitis with a high fatality rate. RNA interference (RNAi) is a powerful tool to silence gene expression and a potential therapy for virus infection. In this study, the antiviral ability of eight shRNA expression plasmids targeting different sites of the NS3 and NS4A genes of JEV was determined in BHK21 cells and mice. The pGP-NS3-3 and pGP-NS4A-4 suppressed 93.9% and 82.0% of JEV mRNA in cells, respectively. The virus titer in cells was reduced approximately 950-fold by pretreating with pGP-NS3-4, and 640-fold by pretreating with pGP-NS4A-4. The results of western blot and immunofluorescence analysis showed JEV E protein and viral load in cells were remarkably inhibited by shRNA expression plasmids. The viral load in brains of mice pretreated with pGP-NS3-4 or pGP-NS4A-4 were reduced approximately 2400-fold and 800-fold, respectively, and the survival rate of mice challenged with JEV were 70% and 50%, respectively. However, the antiviral ability of shRNA expression plasmids was decreased over time. This study indicates that RNAi targeting of the NS3 and NS4A genes of JEV can sufficiently inhibit the replication of JEV in vitro and in vivo, and NS3 and NS4A genes might be potential targets of molecular therapy for JEV infection.

KEYWORDS:

Japanese encephalitis virus; NS3 gene; NS4A gene; RNA interference

PMID:
27593564
DOI:
10.1016/j.gene.2016.08.055
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center