应用正交设计筛选hTERT基因siRNA最佳作用位点

Screening the bestsi RNA targeting hTERT gene by orthogonal design

Abstract: Objective: To screen the best sites and concentration for RNA interference in hTERT gene. Methods: 3 different siRNAs were selected from different positions of the hTERT gene and designed, Synthesized and subcloned into the plasmid pmU6 which including mU6 promoter. The statistical software spss edition 11.5 was used to design orthogonal design, the effect factors were plasmid type, plasmid concentration and time. Their levels are all 3. The plasmids were transfected into the CNE-2Z according to the orthogonal test, and the inhibition of cell proliferation was analyzed by MTT method. The results were confirmed by RT-PCR. Result: The best interfering result were the 0.2ug/uL plasmid pmU6-hTERT-3, which siRNA locates in 2474 to 2489 posintion of hTERT gene on 24 hours after tranfected. Conclusion: It is Simple and convenient to screen the best RNAi sites of the targeting gene according to the orthogonal design.