小干扰RNA对人淀粉样前体蛋白基因的沉默作用(英文)

Silencing effect of amyloid precursor protein triggered by small interfering RNA

目的观察小干扰RNA(siRNA)对人淀粉样前体蛋白(APP)基因的沉默作用。方法该研究将设计好的一对寡核甘酸在细胞内源性地转录成小发夹RNA(shRNA),同时构建了携带两个APP亚型的增强型绿色荧光蛋白(EGFP)的GFP-APP重组质粒。利用EGFP作为报告基因,在COS-7细胞内通过共转染siRNA表达质粒和GFP-APP的重组质粒来观察沉默效应。结果针对APP的siRNA可有效地下调APP基因。结论针对APP的siRNA将不仅在研究APP基因功能和深入研究阿尔茨海默病(AD)的病理分子机制上,而且在运用siRNA这种新型的反义核酸药物治疗AD方面扮演重要角色。

Much needs to be learned about human amyloid precursor protein (APP) gene functions because APP may be etiologically involved in Alzheimer disease(AD). However, physiological function of the APP remains to be elucidated. As a novel technique, RNA interference (RNAi) is quite appropriate to analyze APP gene function. In order to observe silencing effect of small interference RNA (siRNA) directed against APP gene, a designed oligonucleotide was transcribed endogenously into small hairpin RNA (shRNA) and two enhanced green fluorescence protein (EGFP) carried two APP isoforms recombinant plasmids were constructed. It will demonstrate the silencing effect by cotransfection of siRNA expression plasmid and GFP-APP recombinant plasmids into COS-7 cells because EGFP will act as a reporter gene. The siRNA directed against APP could silence APP gene effectively. [Conclusion] Therefore, siRNA directed against APP will play a crucial role in the functional exploration of APP gene.