【摘要】 目的:用RNAi技术抑制哺乳动物神经元中外源报告基因的表达,并探讨该过程中RNAi的时间及剂量效应,为利用RNAi技术研究神经元基因功能提供参考。方法:采用神经细胞来源的细胞系neuro-2a小鼠神经瘤细胞为模型,将外源绿色荧光蛋白(GFP)基因转染到neuro-2a细胞内,利用体外转录法合成siRNA,分为3个剂量组对其进行干扰。结果:Neuro-2a细胞可以被有效地转染,而且siRNA在neuro-2a细胞内可以有效发挥干扰作用。这种干扰作用呈现出一定的剂量效应。结论:RNAi技术可以成功用于neuro-2a细胞抑制基因的表达,这种对GFP表达规律及RNAi剂量效应的研究为利用RNAi技术研究神经元基因功能提供了一定的理论参考和依据。更多还原

【Abstract】 AIM: To assess the effect of RNAi on suppressing the exogenous reporter gene expression in mammalian neurons,and explore the effect of siRNA quantitation on interference efficiency.METHODS: Exogenous green fluorescent protein(GFP) expression vector was transferred into neuro-2a cells,and then the small interference RNA targeting GFP mRNA(siGFP) synthesized by transcription in vitro at three different concentration was used in this experiment.RESULTS: The results showed that the neuro-2a cells can be transfected efficiently and siGFP can inhibit GFP expression greatly.CONCLUSION: RNAi can be applied into mammalian neurons successfully.The research on siRNA quantitation will provide technique support for studying the gene function of neurons in the future.更多还原