RNA干涉技术用于p53基因表达的研究

Study on the expression of tumor suppressor gene p53 by RNAi technology

目的利用RNAi技术研究p53基因表达。方法以质粒为载体,介导体外合成的具有与p53基因目的区域同源的特殊结构的DNA片段,通过脂质体将携带此片段的质粒载体导入COS-7细胞,利用载体的U6-RNA启动子,不断在体内产生21-23ntsiRNA,引起目的基因p53的沉默并用Westernblot检测p53的表达水平。结果通过质粒载体介导体外合成的具有特殊结构的DNA片段,能成功地导致p53蛋白的表达下降。结论RNAi技术可作为研究肿瘤基因的有效手段。

Objective The RNA interference technology was used to research the expression of p53. Methods The DNA temple that is homology with p53 was synthesized in vitro and was mediated by a simple DNA vector into COS-7 cells. Utilizing the well-defined U6-RNA promoter, the vector could efficienty deliver the temple into 21-23 nucleotides short interfering RNA (siR-NA) and the siRNA could induce the homology-dependent degradation of p53. Results The simple DNA vector can successful induce RNAi and efficient and specific knock-down the expression of p53 in COS-7. Conclusion The finding will highlight the general utility of RNAi technology, in future this technology may be become a special method in researching oncogene.