摘要
目的探讨诱导PC12细胞分化的神经细胞靶向沉默Smad7基因特性,同时进行沉默效果检测.方法以Smad7基因为靶目标,设计合成3条siRNA序列,进行细胞转染,利用Real time-PCR和Western blot技术检测沉默效果,筛选出最有效的干扰序列,同时检测出最佳的转染浓度和转染时间.结果针对Smad7基因设计合成及筛选出靶向沉默Smad7基因的干扰序列(siRNA1);siRNA1的最佳转染浓度是4μg/mL;siRNA1的最佳转染时间是24 h;siRNA1对Smad7的抑制效果优于其他干扰序列.结论 siRNA1能有效沉默Smad7基因;lipofectamineTM2000可成功将siRNA1转染至神经细胞,转染效率较高;利用siRNA技术能有效抑制神经细胞.
Objective To detect the targeted silence of Smad7 genes in nerve ischemic cells differentiated from PC12 cells,Method Smad7 genes were taken as the target to design and prepare 3 siRNA sequences for the cell transfection. Real time-PCR and Western blot were used to determinate the mRNA and protein expression of Smad7 by Smad7-siRNA, screened out the most effective interference sequences, and find out the best transfection concentration and the best action time. Results siRNA sequences were prepared and the best targeted interference sequence was Smad7-siRNA. The transfection concentration of Smad7-siRNA was 4 Ixg/mL and the best action time was 24 h. The interfering efficiency of siRNA on Smad7 was better than that of the other sequences. Conclusion Smad7-siRNA can effectively silence Smad7 gene expression. LipofectamineTM 2000 can successfully transfect Smad7-siRNA into nerve cells and the transfection rate was high. The application of siRNA technique can effectively inhibit nerve cells.
出处
《北华大学学报（自然科学版）》
CAS
2014年第2期192-196,共5页
Journal of Beihua University（Natural Science）
基金
吉林省卫生厅基金项目(2013ZC029)
吉林市科技局科研项目(201339033)
