分子生物学第10次课分生-rnai技术.pptxVIP

;Contents;RNAi的发现; petunias Wild type;Su Guo;wild-type embryos;Antisense RNA;C. Elegant worm Wild type ;In 1998: Fire Mello in Nature 证实在RNAi 中，真正起作用的是dsRNA;Sense RNA; Effects of mex-3 RNA interference on levels of the endogenous mRNA (Nomarski DIC micrographs show in situ hybridization of 4-cell stage embryos ) ; 澄清此前一系列令人们困惑的实验现象 揭示一种遗传信息流动的控制机制 RNAi这套重要的基因表达调控机制渐渐地被 揭开其神秘的面纱，为生命科学开拓了 一片新的领域。;Evolution of RNAi;Andrew Z. Fire at 2006 Nobel lectures in Stanford University ;Craig Mello at 2006 Nobel lectures ;From the Nobel Foundations announcement; RNAi occurs in plants, animals, and humans. It is of great importance for the regulation of gene expression, participates in defense against viral infections, and keeps jumping genes under control. ; RNAi is already being widely used in basic science as a method to study the function of genes and it may lead to novel therapies in the future. To learn more about the discovery that will forever change genetics and medicine.; 二. RNAi的作用机制 ( The mechanism of RNAi);The Central Dogma;The Central Dogma; siRNAs (19-23 bp);The guide strand is in the anti-sense orientation with respect to the mRNA.; a large class of evolutionarily conserved. originating from longer transcripts characterized by imperfect hairpin structures.;;; Dicer is a ribonuclease in the RNase III family that cleaves dsRNA and pre-miRNA into small interfering RNA (siRNA) about 19-23 nucleotides long, usually with a two-base overhang on the 3 end. ;2. Effector steps of RNAi;RISC通过碱基配对定位到同源mRNA上，并在 距离siRNA3’端12个碱基的位置切割mRNA。 研究表明每个RISC都包含一个siRNA或miRNA. ;一类庞大的Pr家族，是RISCs的主要成员。 主要含两个结构域：PAZ和PIWI。 PAZ结合到siRNA 的3’端突出硷基； 而PIWI有内切酶活性。 不同的AGO有不同的生物学功能， 如人的AGO2“筹划”了RISCs对于目标mRNA的切割过程；而AGO1 和AGO3则无此功能。 ;Leemor Joshua-Tor, Ph. D. Professor Keck Structural Biology Laboratory Cold Spring Harbor, NY, USA ;王晓东教授 德州大学达拉斯分校生物化学 美国国家科学院中最年轻的院士;(三). Process of RNAi;dsRNA;(四). Pathway of RNAi; Silencing of an endogen