2012年中东爆发了MERS-CoV疫情，逐渐蔓延到欧洲、非洲等地，形势不断恶化，已经引起了全世界极大关注。截至目前，全球已有184例确诊病例，死亡80人。面对日趋严峻的疫情，开发研制安全、有效的疫苗是十分必要的。RNAi由于可以特异性地抑制基因表达, 近年来越来越多的用于抗病毒治疗及预防的研究。在本研究中设计针对MERS-CoV的S基因、N基因的microRNA，首先在细胞水平上初步筛选出有效抑制MERS-CoV复制的microRNA。继而选用腺病毒作为将microRNA安全高效的运输载体，进一步研究重组腺病毒在细胞和恒河猴模型中对MERS-CoV的抑制效果，探讨其在病毒感染免疫保护中的作用。通过本研究的开展，以期获得一种新型抗MERS-CoV制剂，为研究MERS-CoV预防和治疗的新策略提供理论依据。

In 2012, a novel coronavirus, termed MERS-CoV, emerged in the Middle East, and the developing trend has been worsening. To date, 184 human cases of MERS-CoV have been reported with 80 fatalities. Facing the increasing severity of epidemic, it is of utmost importance to develop a safe and effective vaccine against these newly emerging virus. RNA interference (RNAi) refers to the phenomenon of sequence-specific degradation of homologous mRNA induced by double-stranded RNA. It has been successfully utilized to down-regulate endogenous gene expression or suppress the replication of various pathogens in mammalian cells. In this regard, RNAi holds great promise as a novel nucleic acid–based therapeutic against MERS-CoV. In this study, we aimed to develop a novel anti-MERS therapeutic strategies based on RNAi and adenovirus delivery system. We designed and synthesized several artificial mricroRNAs (amiRNAs) complementary to MERS-CoV S and N genes respectively, then cloned these amiRNAs into a commercially available pcDNATM6.2-GW/EmGFP-miR vector. Vero cells were transfected with these plasmids, and then infected with MERS-CoV 24 hours later after transfection, screening for the amiRNAs of inhibiting the replication of MERS-CoV efficiently in cells. For efficient delivery and expression, we cloned these effective amiRNAs into an adenoviral vector, termed Ad5, respectively to evaluate the immune protection in cell and rhesus monkey models. According to this study, we colud got a novel biological agent against MERS-CoV, and provide basic research data for MERS-CoV control.