药学分子生物学：第六章 常用分子生物学技术.pptVIP

* * * * DISCOVERY ? while trying to deepen the purple colour of petunias, researchers introduced a purple pigment-producing gene into the plants. The result ? Plants producing white flowers. Explanation ? the purple-making gene was cosuppressed (turned off) and this phenomenon is called PTGS. The term RNA interference was first used by researchers studying C. Elegans. Injection of the antisense RNA disrupted expression of targetted gene, but ironically, injection of the sense-strand control did too. Explanation ? both antisense and sense RNAs contain dsRNA, which lead to loss of targetted gene expression. Thus, dsRNA is the real trigger of gene silencing. * DISCOVERY ? while trying to deepen the purple colour of petunias, researchers introduced a purple pigment-producing gene into the plants. The result ? Plants producing white flowers. Explanation ? the purple-making gene was cosuppressed (turned off) and this phenomenon is called PTGS. The term RNA interference was first used by researchers studying C. Elegans. Injection of the antisense RNA disrupted expression of targetted gene, but ironically, injection of the sense-strand control did too. Explanation ? both antisense and sense RNAs contain dsRNA, which lead to loss of targetted gene expression. Thus, dsRNA is the real trigger of gene silencing. 3、肿瘤治疗 用RNAi特异性地抑制癌基因、癌相关基因或突变基因的过度表达，使这类基因保持在静默或休眠状态，从而有望用这种新的手段治疗各种恶性肿瘤。 (1)RNAi可应用于敲除点突变激活的癌基因， (2)RNAi应用于抑制插入基因及融合基因表达， (3)RNAi可应用于抑制基因扩增。 3、肿瘤治疗 Survivin基因是迄今为止克隆出的最小凋亡抑制蛋白家族成员，可通过抑制caspase级链反应下游的caspase3，caspase7发挥其抗凋亡作用 。现在研究已证实survivin基因参与了肝癌的发生与发展，并且还与化疗的耐药性相关。降低survivin基因在肝癌细胞中的表达不仅可诱导肝癌细胞的凋亡、抑制肝癌细胞的生长，还可增强其对化疗的敏感性，从而提高肝癌治疗的整体疗效。 3、肿瘤治疗 MDR (Muhidrug resistance)是肿瘤化疗失败的主要原因，形成MDR的最常见的因素是肿瘤细胞MDR基因编码的糖蛋白过度表达，其中，MDR1对于细胞的多重耐药性起着重要的作用。Nieth等表明，特异siRNA可以抑制MDR1基因的表达，从而显著降低肿瘤细胞的耐药性。 4、药物开发 利用RNAi技术来研究药物作用的特异性和机制对于加快药物开发的研制速度大有益处，因为基因组研究成果和高通量的筛选技术，为更好更快发现药靶和候选药物提供了重要基础。在药物开发过程中，用RNAi技术可以对靶基因的功能进行分析，有助于搞清药物的作用机制以及与基因编码产物，及与相应化合物的相互作用，从而更正确地发现药靶，开发更有效的候选药物。 21-2