利用RNA干扰抑制具有多种剪接形式的RNA结合蛋白基因的表达

Inhibition of the Expression of RBPMS Gene by RNA Interference

目的:构建具有多种剪接形式的RNA结合蛋白(RBPMS)基因siRNA的真核表达载体,观察其对RBPMS表达的影响。方法:利用RNA干扰(RNAi)技术,设计并合成了2条针对RBPMS基因的siRNA,将其克隆到siRNA表达载体pSliencer2.1-U6neo上。将重组质粒和带FLAG标签的RBPMS共转染293T人胚肾细胞,通过Western印迹检验RNAi效应。结果:测序证明成功构建了RBPMSsiRNA真核表达载体;Western印迹表明构建的siRNA能有效地抑制RBPMS基因的表达。结论:构建了RBPMSsiRNA的真核表达载体,该siRNA能有效地抑制RBPMS基因的表达。

Objective： To construct the eukaryotic expression vector of RNA binding protein with multiple splicing （RBPMS） siRNA and investigate the effect of RBPMS siRNA on the expression of RBPMS gene. Methods： Two RBPMS siRNAs were designed by RNA interference（RNAi） technique and the RBPMS siRNAs were inserted into pSliencer 2.1-U6 neo expression vector. Then human embryo kidney 293T cells were cotransfected with the recombination plasmids and FLAG-tagged RBPMS expression vector. The effect of RBPMS siRNAs on the expression of RBPMS gene was identified by Western blot. Results： The expression vectors of RBPMS siRNAs were constructed and confirmed by DNA sequencing. Western blot showed that RBPMS siRNAs could effectively inhibit the expression of RBPMS gene. Conclusion： The eu- karyotic expression vectors of RBPMS siRNAs were constructed successfully. The siRNAs could effectively inhibit the expression of RBPMS gene.