Construction of Short Hairpin RNA Vector with σNS ＆ σC Genes of Avian Reovirus and Determination of Interference Effect

[Objective] The aim was to explore novel method for treatment of Avian Reovirus. [Method] According to the design principle of siRNA target sequences, siRNA templates were designed and synthesized and then cloned into the shRNA expression vector, namely, pSilencer-CMV 4.1 neo. Short hairpin RNA vector C1, C2, C3, which contain σC gene, and shRNA vector NS1, NS2, NS3, which contain σNS gene, were constructed separately. The constructed shRNA vectors and negative control were co-transfected into DF-1 cells with the eukaryotic expression vector pEGFP-σC and pEGFP-σNS, respectively. [Result] Observation through fluorescence microscope indicated that the constructed 6 shRNA could inhibit the expression of fusion protein to different degrees. In addition, results of Real-time PCR suggested that C3 and NS1 have the best interference effect to the viral duplication in vitro. [Conclusion] Construction and selection of specific shRNA expression vectors inhibiting Avian Reovirus are significant for researching effects of σC and σNS proteins in infection and duplication of ARV, providing new idea for ARV antiviral therapy.