RNA干扰用siRNA的体外转录合成

Synthesis of siRNA by in vitro for Transcription RNA Interference

RNA干扰(RNAinterference,RNAi)作为一种特异性沉默基因表达的方法,正在成为研究基因功能、胚胎发育及病毒性疾病治疗的重要工具,而获得符合干扰要求的短双链干扰RNA(smallinterferenceRNA,siRNA)是进行RNA;研究的首要步骤。本研究建立了体外转录合成siRNA方法,并用其生成的siRNA干扰鸡成纤维细胞外源绿荧光蛋白(GFP)基因和内源3-磷酸甘油醛脱氢酶(GAPDH)基因的表达。结果显示,siRNA能特异性降低鸡成纤维细胞中内外源基因的表达。本实验认为这种以DNA为模板体外转录合成siRNA方法操作简单、成本低、产物得率高,值得从事RNA研究者参考借鉴。

RNA interference (RNAi) has developed into a powerful tool for studying gene function, embryonic development and virosis therapy. The first step of RNAi is to acquire the small interference RNA(siRNA). In this study , the siRNAs were synthesized by the approach of in vitro transcription . They were used to interfere the expression of green fluorescent protein (GFP) exterior gene and Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) inner gene in chicken fibroblast cell. The results indicate that the synthetic siRNAs can specifically decrease the expression of GFP gene and GAPDH gene. This approach using a DNA template to synthesize small RNA in vitro transcription is worthy of reference for simple operation, low cost and high productivity.