RNAi干涉技术及其应.ppt

RNAi干涉技术及其应用 RNA Interference (RNAi) technique and its application 田生礼 生命科学学院 RNA单链线型分子，自身形成二级或三级结构。除tRNA外，几乎都与蛋白质结合。 功能：信息分子，功能分子。 重要功能：①参与蛋白质合成； ②具有生物催化剂的功能 ③参与基因的表达调控 ④与生物体的进化有关 核糖体，信息体，信号识别颗粒，剪接体，编辑体等。 什么是RNAi？ RNAi（RNA interference）是指在生物体细胞内，dsRNA引起同源mRNA的特异性降解，因而抑制相应基因表达的过程。 一种转录后水平的基因沉默 生物体内普遍存在 抵御外在感染的重要保护机制 RNAi的发展历程 1995，RNAi现象首次在线虫中发现。Cell，1995 ，81：611-620 1998，RNAi概念的首次提出。Nature, 1998, 391（6669）：806 RNA interference. Degradation of mRNA molecules specified by complementary double-stranded RNA 1999，RNAi作用机制模型的提出。Cell，2000，101（1）：25-33 在线虫、果蝇、拟南芥及斑马鱼等多种生物内发现 RNAi现象。 2001，RNAi技术成功诱导培养的哺乳动物细胞基因沉 默现象。Nature，2001，411（6836）：494－498 RNAi技术被《Science》评为2001年度的十 大科技进展之一。 2002至今，有关RNAi的研究蓬勃开展，是分子生物学领域最为热门的方向之一。 Five parts Background Theory Technology Application Problems Summary Figure 2. SOAKING C. elegans WORKS ALMOST AS EFFECTIVELY AS INJECTING. demonstrating the results of mex-3 in situ hybridization following an RNAi soaking protocol. The left panels show the wildtype pattern of endogenous mex-3 mRNA in untreated adults and embryos. The right panels show loss of mex-3 staining following soaking in mex-3 dsRNA. Endogenous mex-3 RNA is greatly reduced. Soaking as a delivery method does NOT work for many other nematode species. Images by Jeff Norman (in 1999), Another research group who were expressing genes from the potato virus in tobacco plants, hoped that viral proteins produced by the plants would stimulate a defense mechanism allowing the plants to resist subsequent attack by the virus. To their surprise, the plants with the strongest resistance to the virus were those in which the introduced gene was silent[9]. Using C. elegans, it was demonstrated that injection of double stranded RNA was more effective in gene silencing than injection of sense