【摘要】 [目的]研究3种不同长度的四联shRNA载体及其在细胞与个体水平的沉默效应差异。[方法]以EGFP为靶标基因,用hU6、mU6、h7SK、hH1 4种启动子,构建21、27、29 bp的3种四联沉默载体,通过转染Vero细胞和注射小鼠肌肉,以实时荧光定量PCR的方法检测绿色荧光蛋白基因的mRNA水平。[结果]3种载体都有很强的沉默效应,其在细胞水平上的沉默效应远高于个体水平上,可以通过未成年鼠的肌肉注射进行转基因。[结论]多位点串联表达shRNA是一种可以达到较好干扰效果的方法,肌肉注射将是一个简便的转基因途径。更多还原

【Abstract】 [Objective] The aim was to compare the silencing effect of three vectors containing different lengths of shRNA quartet at cellular level and individual level.[Result] To silence EGFP gene,three shRNA quartets of 21,27 and 29 bp formed by hU6,mU6,h7SK and hH1 promoters were constructed and then cloned into silent vectors.After the vectors were transfected into Vero cells or injected into mouse muscle,real-time fluorescence quantitative PCR was conducted to detect the mRNA level of EGFP gene.[Result] All the three vectors showed strong silence effect;in which,the silencing effect at the cellular level was far higher than that at individual level.[Conclusion] Tandem shRNA expression vector may show better silencing effect;intramuscular injection maybe a transgenic method for baby mouse.更多还原