论文部分内容阅读
目的制备靶向CDC42基因的小干扰RNA分子(siRNA)并检测其对CDC42基因表达的抑制效果。方法针对CDC42基因的不同区域设计4条siRNA分子,体外转录法合成siRNA分子,将siRNA与CDC42基因真核表达载体共转染HEK293T细胞,Westernblot法测定CDC42蛋白含量。结果合成了siR1、siR2、siR3、siR4共4条siRNA分子,siR3对CDC42基因表达的抑制率为95%,siR1、siR2以及siR4对CDC42基因表达的抑制作用不明显。siR3对HEK293T细胞生长无明显影响。结论体外转录法制备的siR3分子能够高效特异地抑制CDC42基因表达。
Objective To prepare small interfering RNA (siRNA) targeting CDC42 gene and test its inhibitory effect on CDC42 gene expression. Methods Four siRNA molecules were designed according to different regions of CDC42 gene. SiRNA molecules were synthesized by in vitro transcription. The siRNA and CDC42 eukaryotic expression vector were co-transfected into HEK293T cells. The protein content of CDC42 was determined by Western blot. Results A total of 4 siR1, siR2, siR3 and siR4 siRNA molecules were synthesized. The inhibitory rate of siR3 on CDC42 gene expression was 95%. The inhibitory effect of siR1, siR2 and siR4 on CDC42 gene expression was not obvious. siR3 had no significant effect on the growth of HEK293T cells. Conclusion In vitro transcription of siR3 molecules can efficiently and specifically inhibit CDC42 gene expression.