分子生物学专题一----RNAi-1幻灯片.pptVIP

Contents RNAi involves in viral defense、transposon silencing and gene regulation. Gene silencing Contents 1995-C. elegans 1998-C. elegans 1998-C. elegans Contents 3. Amplification of Silencing Contents Two methods siRNA expression vector siRNA expression vector siRNA expression vector siRNA expression vector Contents Applications of RNAi Gene function Gene therapy ---Diseases by virus ---Diseases by tumor In 2002, Jacque designed siRNAs against LTR, Vif and Nef of the HIV genome.The expression of genes is decreased by 95%. In 2010, Saki blocked the expression of CCR5, and the mice showed a stable resistance to HIV. To Design a kind of dsRNA can block the expression of some homologous genes. Reading Potent and speci?c genetic interference by double-stranded RNA in Caenorhabditis elegans. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC.(1998) Applications of RNA Interference in Mammalian Systems. Martin SE et al. Annu Rev Genomics Hum Genet. (2007) THE SILENT REVOLUTION: RNA Interference as Basic Biology, Research Tool, and Therapeutic. Dykxhoorn DM, Lieberman J.Annu Rev Med. (2005) 思考题 1.比较siRNA和miRNA的异同点。 2.在Fire等的Nature文章中，他们是怎样想到用dsRNA的？ Entrez PubMed Search Terms: RNAi Number of Publications RNAi of C. elegans GFP-----green fluorescence protein (绿色荧光蛋白) or GFP dsRNA In 2001 and 2002, RNAi was named by science magazine as one of the ten major breakthroughs in Natural Science. Andrew Z. Fire Craig C. Mello How does RNAi work? Theory Definition Discovery Design Application Initiation Execution Amplification General mechanism of RNAi short-interfering RNA Dicer 1.Initiation（起始阶段） The dsRNA cleavage by Dicer generating siRNAs. Theory Dicers belong to Class III of RNase III, a family of endo-ribonucleases that show specificity for double-stranded RNA (dsRNA)----PAZ domain, helicase domain plus two RNase III domains Theory （belong to Class III of RNase III） Dicer Model for Dicer catalysis The PAZ domain bind