RNA干扰对Smad7基因的抑制作用

Inhibitory Effect of RNA Interference on Smad7 Gene Expression

小分子干扰RNA(siRNAs)可以高效、特异地阻断体内同源基因的表达,促进同源mRNA降解,称为RNA干扰(RNAi)。本研究旨在探讨Smad7基因的siRNAs是否能抑制基因的表达。利用RNA干扰技术,设计并合成了针对Smad7基因的siRNAs,用脂质体转染法瞬时转染BEP2D和BERP35T2细胞,用Northernblot法检测RNAi效应;同时设计并合成了绿色荧光蛋白(GFP)的siRNAs,瞬时转染稳定表达绿色荧光蛋白的BERP35T2细胞,检测荧光强度有无改变。结果表明RNA干扰技术能明显抑制Smad7基因的表达,并能显著减弱绿色荧光的表达强度,为进一步研究Smad7基因功能及TGF-β信号转导通路奠定了基础。

The tiny RNAs are termed 'short interfering RNAs'(siRNAs) depending on their origin. They down-regulate gene expression effectively by binding to complementary mRNAs and triggering mRNAs degradation(RNAi). The current research was designed to investigate whether siRNAs can inhibit the expression of Smad7 gene. Gene transfection and Northern blot were performed to determine the effect of gene inhibition of Smad7 by RNA interference in BEP2D and BERP35T2 cells. Meanwhile, fluoroscopy was used to evaluate impact of GFP siRNAs on GFP expression. It could be conclude that RNAi can inhibit specific expression of Smad7 gene and GFP gene. The results lay foundation for further study on function of Smad7 gene and TGF-β signal transduction pathway.