title: Inhibition of highly pathogenic PRRSV replication in MARC-145 cells by artificial microRNAs.
abstract: BACKGROUND
  Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) has caused large economic losses in swine industry in recent years. However, current antiviral strategy could not effectively prevent and control this disease. In this research, five artificial microRNAs (amiRNAs) respectively targeted towards ORF5 (amirGP5-243, -370) and ORF6 (amirM-82, -217,-263) were designed and incorporated into a miRNA-based vector that mimics the backbone of murine miR-155 and permits high expression of amiRNAs in a GFP fused form mediated by RNA Pol II promoter CMV.
  
  
  RESULTS
  It was found that amirGP5-370 could effectively inhibit H-PRRSV replication. The amirM-263-M-263, which was a dual pre-amiRNA expression cassette where two amirM-263s were chained, showed stronger virus inhibitory effects than single amirM-263. H-PRRSV replication was inhibited up to 120 hours in the MARC-145 cells which were stably transduced by recombinant lentiviruses (Lenti-amirGP5-370, -amirM-263-M-263). Additionally, efficacious dose of amirGP5-370 and amirM-263 expression did not trigger the innate interferon response.
  
  
  CONCLUSIONS
  Our study is the first attempt to suppress H-PRRSV replication in MARC-145 cells through vector-based and lentiviral mediated amiRNAs targeting GP5 or M proteins coding sequences of PRRSV, which indicated that artificial microRNAs and recombinant lentiviruses might be applied to be a new potent anti-PRRSV strategy.
authors: State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, P. R. China.
authors: Shuqi
authors: S
authors: Xiao
authors: Qiwei
authors: Q
authors: Wang
authors: Jintao
authors: J
authors: Gao
authors: Liangliang
authors: L
authors: Wang
authors: Zuyong
authors: Z
authors: He
authors: Delin
authors: D
authors: Mo
authors: Xiaohong
authors: X
authors: Liu
authors: Yaosheng
authors: Y
authors: Chen
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001688:Biological Products / Q000494:pharmacology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D035683:MicroRNAs / Q000494:pharmacology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000187:drug effects* / Q000254:growth & development*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D001688:Biological Products; D035683:MicroRNAs