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

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。

Inhibition of the Expression of RBPMS Gene by RNA Interference

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 eukaryotic expression vectors of RBPMS siRNAs were constructed successfully.The siRNAs could effectively inhibit the expression of RBPMS gene.