应用实时荧光定量PCR筛选TRF2 siRNA最佳靶序列

SCREENING OF EFFECTUAL SIRNA-MEDIATED TRF2 BY REAL-TIME FLUORESCENCE QUANTITATIVE PCR

有效siRNA的筛选是RNAi研究的关键点之一。[目的]筛选有效的TRF2 siRNA,为应用RNAi技术抗TRF2研究提供实验基础。[方法]采用化学合成法合成3段针对TRF2的siRNA、应用脂质体LipofectamineTM2000转染试剂将3段siRNA转染人乳腺癌MCF-7细胞、实时荧光定量RT-PCR技术检测TRF2 mRNA表达水平以确定干扰效果。[结果]3段TRF2 siRNA中有2段siRNA可有效降低TRF2 mRNA表达。[结论]利用化学合成法合成siRNA,应用实时荧光定量PCR技术检测目的基因干扰效果可用于快速、高效筛选特异性基因表达抑制的siRNA。

[Objective] To screen effectual siRNA-mediated TRF2, and lay the foundation for further studying on anti- TRF2 by RNA interfere. [Methods] Three TRF2-specific double stranded siRNAs were designed targeting different regions of TRF2 mRNA by chemic method, siRNAs were electroporated into MCF-7 ceils by LipofectamineTM2000, TRF2 mRNA expression level were determined by Real-time fluorescence quantitative RT-PCR. [ Results ] Two of the three customized TRF2-siR- NAs could effectively inhibit the expression 9f TRF2 mRNA. [Conclusion] siRNA synthesizing with chemic method and determining gene expression with Real-time fluorescence quantitative RT-PCR could rapidly, specifically and effectively screen siRNA which could suppress the expression of TRF2 mRNA.