Suppression of porcine reproductive and respiratory syndrome virus replication in MARC-145 cells by shRNA targeting ORF1 region

Abstract

Porcine reproductive and respiratory syndrome (PRRS) is an economically important disease in swine producing area. The current vaccine strategies cannot provide complete protection against PRRSV. The objective of this study was to determine if specific short-hairpin RNA (shRNA) directed against different genomic regions of ORF1b of PRRSV could be utilized to inhibit virus replication in MARC-145 cells. Two shRNA expression vectors targeting ORF1b gene of PRRSV were constructed and delivered into MARC-145 cells, and then infected with PRRSV. The results showed that PRRSV-specific cytopathic effect (CPE) could be inhibited in the cells transfected with pSUPER-P2 and pSUPER-P3, and the virus titers in the cells transfected with pSUPER-P2 and pSUPER-P3 were lower than those control cells by approximately 100 fold. Moreover, the expression of ORF1 of PRRSV in the cells was reduced both at RNA and protein levels comparing to the controls. It indicated that vector-based shRNA targeting ORF1 region could effectively inhibit PRRSV replication in MARC-145 cells.

Glycyrrhizic‐Acid‐Based Carbon Dots with High Antiviral Activity by Multisite Inhibition Mechanisms

“…Previous studies have investigated many microbicides as potential anti‐PRRSV agents to eliminate PRRSV in the body at any stage of PRRSV presence. These microbicides include protein,35 microRNAs,36,37 antisense oligonucleotides,38–41 chemical compounds,30,42 and nanobodies 43. They are shown to inhibit virus adsorption, invasion, replication, release, and cell‐to‐cell spread by restricting cell tropism and targeting the PRRSV genome.…”

“…They are shown to inhibit virus adsorption, invasion, replication, release, and cell‐to‐cell spread by restricting cell tropism and targeting the PRRSV genome. However, they have a relatively slight anti‐PRRSV effect, with a maximal inhibitory effect of ≈3 orders of magnitude 32,36–41,43–47…”

Mir-331-3p Inhibits PRRSV-2 Replication and Lung Injury by Targeting PRRSV-2 ORF1b and Porcine TNF-α

“…Li et al (41) showed that PRRSV-specific cytopathic effect (CPE) could be inhibited in the cells by shRNA targeting ORF1b, and that cellular virus titers were decreased by ∼100-folds compared with those of control cells. Li et al (42) reported that two recombinant adenoviruses expressing shRNA could effectively inhibit PRRSV replication in vitro and in vivo by targeting ORF1b of PRRSV.…”

“…Of them, Nsp9 and Nsp10 were key enzymes for RNA synthesis of arterial virus, and closely related to the replication efficiency in vitro and in vivo and related to the increased pathogenicity and fatal virulence for piglets ( 12 ). Li et al ( 41 ) showed that PRRSV-specific cytopathic effect (CPE) could be inhibited in the cells by shRNA targeting ORF1b , and that cellular virus titers were decreased by ~100-folds compared with those of control cells. Li et al ( 42 ) reported that two recombinant adenoviruses expressing shRNA could effectively inhibit PRRSV replication in vitro and in vivo by targeting ORF1b of PRRSV.…”

Protective efficacy of adenovirus-mediated small interfering RNAs against encephalomyocarditis virus challenge in mice

“…Because of the high rapidity and specificity of the RNAi effect, it represents a new feasible approach for developing effective antiviral treatments. And it has been employed to induce antiviral effects against various viruses, including porcine circovirus type 2 (PCV-2) (Feng et al, 2008), foot-and-mouth disease virus (Chen et al, 2006), porcine reproductive and respiratory syndrome virus (Huang et al, 2006;Li et al, 2007), influenza A virus (Zhou et al, 2007), human immunodeficiency virus Huelsmann et al, 2006), and herpes simplex virus (Muylaert and Elias, 2007).…”