Inhibition of highly pathogenic PRRSV replication in MARC-145 cells by artificial microRNAs

Virology Journal

Yaosheng Chen¹  Xiaohong Liu¹  Delin Mo¹  Zuyong He¹  Liangliang Wang¹  Jintao Gao¹  Qiwei Wang¹  Shuqi Xiao¹ 

[1] Pig Improving and Breeding Project Technology Research Exploitation Center of Guangdong, Guangzhou 510006, P. R. China

关键词: Lentivirus;    artificial miRNA;    RNAi;    Highly pathogenic PRRSV;   

Others  :  1155759
DOI  :  10.1186/1743-422X-8-491

 received in 2011-09-01, accepted in 2011-11-01,  发布年份 2011

【 摘 要 】

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.

【 授权许可】

   
2011 Xiao et al; licensee BioMed Central Ltd.

【 图 表 】

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