RNA干扰对阿尔茨海默病家族性瑞典型APP突变的沉默作用邱

Induction of silencing effect of Alzheimer disease familial Swedish mutant APP by RNA interference

目的诱导小干涉RNA (small interference RNA, siRNA )对阿尔茨海默病淀粉样前体蛋白家族性瑞典型突变(APPswe)的沉默作用。方法设计一对针对APPswe的寡核苷酸,构建成内源性表达相应siRNA的质粒,与构建的APP-EGFP重组质粒瞬时共转染COS-7细胞,利用GFP报告基因来间接观察siRNA对APPswe的沉默作用。结果siRNA可有效地下调突变型APP,相比APPwt而言,针对突变型APP的siRNA对APPswe的沉默作用更强。结论针对APPswe的siRNA可有效地沉默APPswe基因,并有一定的等位基因特异性。因此,针对APPswe的siRNA将在阿尔茨海默病治疗研究领域扮演重要角色。

Objectives To observe the silencing effect of amyloid precursor protein (APP) carrying Swedish mutation of familial Alzheimer disease triggered by small interference RNA (siRNA). Methods We designed one oligonucleotide against Swedish mutant APP(APPswe) which was endogenously expressed by small interference RNA(siRNA) expressing plasmid . In order to observe the silence effect of siRNA on APP, COS-7 cells were transiently co-transfected with APP-EGFP recombinant plasmid and siRNA expressing plasmid. The silencing effect of siRNA was determined by the assay of reporter gene-GFP expression levels. Results APPswe expression in COS-7 cells was knocked down by the application of siRNA. APPswesiRNA was more effective in silencing APPswegene than wild type APP (APPwt) gene. Conclusion APPswe siRNA can effectively silence APPswe gene expression and such effect is allele-specific. Therefore, siRNA directed against APPswe may have therapeutic potential for the treatment of Alzheimer disease.