Inhibition of porcine reproductive and respiratory syndrome virus by specific siRNA targeting Nsp9 gene

Infect Genet Evol. 2014 Dec:28:64-70. doi: 10.1016/j.meegid.2014.08.008. Epub 2014 Aug 20.

Abstract

To screen siRNAs for effectively inhibiting the replication of porcine reproductive and respiratory syndrome virus (PRRSV). Four pairs of siRNA targeting Nsp9 gene of PRRSV and one non-efficient pair used as control were designed, synthesized and cloned into pSilencer4.1-CMV neo, designated as pSi-294, pSi-367, pSi-409, pSi-1488, pSi-Ctr. The recombinant plasmids were transfected into Marc-145 cells and infected with PRRSV 24h post transfection. Subsequently, IFA, real-time PCR, TCID50 and western blot were used for evaluating the inhibitory effect of the siRNA. IFA and western-blot results showed that pSi-294, pSi-1488 can effectively inhibit the expression of Nsp9 and M protein of PRRSV, real-time PCR result showed that the expression of Nsp9 gene were decreased from 86.56% to 93.66% compared to the negative control. siRNAs can be used as candidates for basic research of PRRSV.

Keywords: Nsp9; PRRSV; RNAi.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytopathogenic Effect, Viral
  • Gene Expression Regulation, Viral
  • Genes, Viral*
  • Models, Molecular
  • Porcine respiratory and reproductive syndrome virus / genetics*
  • Protein Conformation
  • RNA Interference*
  • RNA, Small Interfering / genetics*
  • Viral Nonstructural Proteins / chemistry
  • Viral Nonstructural Proteins / genetics
  • Virus Replication / genetics*

Substances

  • RNA, Small Interfering
  • Viral Nonstructural Proteins