RNA干扰抑制ST细胞Ⅰ型干扰素受体-2表达的研究

Study on Expression Inhibition of TypeⅠInterferon Receptor Gene-2 in ST Cells with RNA Interference

应用RNA干扰技术建立猪睾丸(ST)细胞Ⅰ型干扰素受体-2(IFNAR-2)基因knock down模型。采用FuGENE HD转染剂将pSiCMVE1、pSiCMVE2和pSiCMVE3干扰载体分别转染ST细胞,经荧光定量PCR和Western blot分别检测IFNAR2mRNA和蛋白水平表达变化。结果显示,与对照组比,转染干扰载体组均可明显降低IFNAR2 mRNA和蛋白的表达(P<0.05)。RNA干扰技术能够有效抑制IFNAR2基因的表达,为进一步研究IFNAR2基因的生物学功能提供可靠的手段,为研究IFNAR2在病毒感染中的作用奠定基础。

To establish a type I interferon receptor gene-2（IFNAR2） knock down model of ST cells with RNA interference technique,three interfering vectors of pSilencer-4.1, named pSiCMVE1, pSiCMVE2 and pSiCMVE3 were constructed and transfected into ST cells by FuGENE HD. The expression changes of IFNAR2 mRNA and IFNAR2 protein in ST cells were evaluated with qPCR and Western blot respectively. The results showed that compared with normal control, the expressions of IFNAR2 mRNA and protein were significantly reduced by siRNA transfection in ST cells（P〈0.05）. RNA interference technique can effectively down regulate the expression of IFNAR2 gene in ST cells specifically. This study provided a novel and reliable method for further study on the biological functions and the role of IFNAR2 in virus infection.