Inhibition of encephalomyocarditis virus replication by shRNA targeting 1C and 2A genes in vitro and in vivo

Vet Microbiol. 2020 May:244:108664. doi: 10.1016/j.vetmic.2020.108664. Epub 2020 Apr 2.

Abstract

Encephalomyocarditis virus (EMCV) infects many mammalian species, causing myocarditis, encephalitis and reproductive disorders. The small interference RNA (siRNA) targeting to the virus has not been understood completely. Here, two out of six interference sequences were screened to inhibit significantly EMCV replication by using recombinant plasmids expressing small hairpin RNA (shRNA) targeting to the viral 1C or 2A genes in BHK-21 cells. And two recombinant adenoviruses expressing the shRNAs were constructed and named as rAd-1C-1 and rAd-2A-3. They inhibit EMCV replication in BHK-21 cells in protein levels, as well as the virus yields by approximately 1000 times. Furthermore, they provide high protective efficacy against the challenge with virulent EMCV NJ08 strain in mice. And the EMCV loads in the live mice in rAd-1C-1 and rAd-2A-3 groups decrease by more than 90 % compared with those in the dead mice in the challenge control groups at the same times. It indicates that the adenoviruses medicated shRNA targeting to 1C and 2A genes might provide a potential strategy for combating EMCV infection.

Keywords: 1C gene; 2A gene; EMCV; Short hairpin RNA (shRNA).

MeSH terms

  • Adenoviridae / genetics
  • Animals
  • Cell Line
  • Encephalomyocarditis virus / genetics*
  • Encephalomyocarditis virus / physiology
  • Female
  • Genes, Viral*
  • Genetic Vectors
  • Mice
  • Mice, Inbred BALB C
  • RNA Interference*
  • RNA, Small Interfering / genetics
  • Viral Load
  • Virus Replication / genetics*

Substances

  • RNA, Small Interfering