武汉大学生命科学学院：《分子生物学》（英文版）Part IV：Regulation CHAPTER18 The mechanism of RNAi and the role of miRNA in development and cancer

Molecular biology of the Gene 5/E - Watson et al. ( 2004) Part l: chemistry and genetics Part l: Maintenance of the Genome Part l: Expression of the Genome Part Iv: Regulation Part v: Methods 2005-5-10 刻博来字 Wuhan Universit

2 Molecular Biology of the Gene, 5/E --- Watson et al. (2004) Part I: Chemistry and Genetics Part II: Maintenance of the Genome Part III: Expression of the Genome Part IV: Regulation Part V: Methods 2005-5-10

Chapter 16 Regulation principles and How genes are regulated in bacteria Chapter 17 Basic mechanism of gene expression in eukaryotes Chapter 18 The mechanism of rnai and the role of mirna in development and cancer

3 Chapter 16 Regulation principles and How genes are regulated in bacteria Chapter 17 Basic mechanism of gene expression in eukaryotes Chapter 18 The mechanism of RNAi and the role of miRNA in development and cancer

Molecular Biology Course Chapter 8: RNA in requlation The mechanism of rnai and the role of miRNA in development and cancer 隅来字 Wuhan University

4 Chapter 8: RNA in regulation The mechanism of RNAi and the role of miRNA in development and cancer •Molecular Biology Course

CHAPTER 18 RNAi and mia in development and cancergenesis 一、RNA干扰及其机制 Topic 1: RNA interference and its mechanism 劣溪字 Wuhan University

5 Topic 1: RNA interference and its mechanism CHAPTER 18 RNAi and miRNA in development and cancergenesis 一、RNA干扰及其机制

1 Double-stranded rNa inhibits expression of genes homologous to that rNA. [ phenomena-现象] 双链RNA抑制含其同源序列基因的表达

6 1 Double-stranded RNA inhibits expression of genes homologous to that RNA. [phenomena-现象] 双链RNA抑制含其同源序列基因的表达

2006年的诺贝尔生理学奖获得者: The nobel prize in physiology or Andrew 2。Fre Medicine 2006 Craig C. Mello for their discovery of RNA interference- gene silencing by double stranded RNA Potent and specific genetic interference by double-stranded rna in Caenorhabditis elegans Andrew Fire, SiQun Xu, Mary K Montgomery, Steven A Kostas*t, Samuel E Driver& Craig C. Mello: Photo: L Cicero/stanford Photo: R. Carlin/UMMAS Andrew Z Fire Craig C. Mello NATURE VOL 39119 FEBRUARY 1998 41/2 of the prize 4 1/2 of the prize USA USA Stanford University School University of of Medicine Massachusetts Medical Stanford CA USA School Worcester, MA, USA

7 2006年的诺贝尔生理学奖获得者： Andrew Z. Fire Craig C. Mello

Fig 2. Analysis of RNA-interference effects in individual cells, Fluorescence micrographs show progeny of injected animals from GFP-reporter strain PD4251(a C elegans strain expressing GFP fluorescence protein)(使用外源导入的报告基因) Control dsrNA ds-gfp RNA young larva(幼虫) Adu(成虫)选 adult body wall at high magnification 高放大倍数的 成虫体壁) NATURE VOL 39119 FEBRUARY 19988

8 Fig 2. Analysis of RNA-interference effects in individual cells. Fluorescence micrographs show progeny of injected animals from GFP-reporter strain PD4251 (a C. elegans strain expressing GFP fluorescence protein) (使用外源导入的报告基因). Young larva (幼虫) Adult (成虫) adult body wall at high magnification (高放大倍数的 成虫体壁) Control dsRNA ds-gfp RNA

Fig 3. Effects of mex-3 RNA interference on levels of the endogenous RNA(in situ hybridization in embryos)(胚胎的原位杂交) No hybridization hybridization and staining B (endogenous mex-3 RNA) +antisense +ds mex-3 RNA C +hybridization D hybridization 998Q

9 Fig 3. Effects of mex-3 RNA interference on levels of the endogenous mRNA (in situ hybridization in embryos) (胚胎的原位杂交). adult body wall at No hybridization and staining + hybridization (endogenous mex-3 RNA) +antisense +hybridization +ds mex-3 RNA +hybridization

An plant immune system: Virus-induced gene silencing(植物病毒引起的基因沉默) Most plant viruses have single-stranded RNA genomes, which are released from the protein coat of their virus particles as they enter a cell. Their genomic RNA is then replicated by the virus encoded rna dependent rna polymerase to produce sense and antisense RNA, which can hybridize to form dsRna and trigger an RNAi response against their own sequences 10

10 An plant immune system: Virus-induced gene silencing (植物病毒引起的基因沉默). Most plant viruses have single-stranded RNA genomes, which are released from the protein coat of their virus particles as they enter a cell.Their genomic RNA is then replicated by the virus encoded RNA￾dependent RNA polymerase to produce sense and antisense RNA, which can hybridize to form dsRNA and trigger an RNAi response against their own sequences

2. Short interfering RNa (siRNAs )are produced from dsRNa and direct machinery that switch off genes in various Way.[ Mechanism机制] 从双链RNA产生的小干扰RNA可以指 导用不同机制关闭基因的细胞机器

11 2. Short interfering RNA (siRNAs) are produced from dsRNA and direct machinery that switch off genes in various way. [Mechanism-机制] 从双链RNA产生的小干扰RNA可以指 导用不同机制关闭基因的细胞机器