Nobel Prize 2006

  • View

  • Download

Embed Size (px)


Nobel Prize 2006. RNA interference - gene silencing by double-stranded RNA. 遺傳中心法則. Andrew Z. Fire Craig C. Mello. 1986 年始進行「線蟲早期發展的基因調控」的研究→和線蟲結下不解之緣 直至 2003 年, Fire ,一直以線蟲為模型,從事遺傳變異影響細胞和生物體的相關研究。 04 年獲選為美國科學院院士. 1982 年加入 David Hirsh 的實驗室並在那奠下分子生物學之基礎 - PowerPoint PPT Presentation

Text of Nobel Prize 2006

  • Nobel Prize 2006RNA interference - gene silencing by double-stranded RNA

  • Andrew Z. Fire Craig C. Mello 19862003Fire04 1982David Hirsh 2005

  • Pigmented petunia experiment,1990Chalcone synthase (CHS) They attempted to overexpress chalcone synthase (CHS)Unexpected resultsForty-two percent of plants presented abnormal.Unclear mechanism

  • Virus-induced recovery N.clevelandii plantTomato black ring nepovirus strain W22 Primary and secondary inoculationRecovery

  • Resistance in the recovered leaves is specific for viruses that have RNA sequences that are similar to the virus used for primary inoculation.

  • 1998Caenorhabditis elegans()C.elegans

  • RNAi

  • Sense RNARNAAntisense RNASense RNARNAUnc-22 gene

  • 1.unc-22 geneC. elegans

  • GFP green fluorescent protein

    gfp-gene GFP

  • UNC-22geneGFP geneLacZL gene

  • GFPds-Unc22RNAds-lacZL RNAds-gfpG RNA

  • mex3antisense mex-3 RNAds mex-3RNA

  • mRNAAntisense oligonucleotideRibozymeRNAimiRNAdsRNAsiRNA

  • Ribozyme

  • Antisense oligonucleotide

  • Control of RNAi and gene expressiondsRNA Affection of virusTransposonsExogenous RNAsiRNAdsRNA miRNA

  • DicerRNase III domains -cleave dsRNA or pre-miRNAconnector helix -dsRNAplatform domain -amino-terminal helicase domain -RNasePAZ domain -dsRNA 3

  • RISC (RNA-induced silencing complex)PAZ domain-from dicerN domain-connect PAZ and PIWIPIWI domain- degrade mRNAMid domain-siRNA 5 mRNA 3

  • siRNA binds with RISCDicer cleaves dsRNARNA interference-siRNAdsRNA is cleaved into small fragmentsmRNA is degraded

  • siRNAdicerRISCDcr2dicerplatform domainhelicase ATP-binding domainATPconnector helixdsRNARISCanti-sensesiRNA

  • miRNA

  • c-Myc-regulated microRNAs miR-17~miR-92

  • RNAi


  • RNAi applicationThe source of siRNA1.synthesize in test tube PCR siRNA target cells2.synthesize in organism

    siRNA target cellsreplication

  • Design a siRNA expression vector

  • Design a siRNA sequenceChoose proper siRNA expression vector

    5~6 A(T) tail

    Select different parts of sequence of several target genes Ex: AUCCGAAGCCUACUCCUUAU

  • RNAi

    2003AcuityRNAimacular degenerationSirnaRNAiHBV95RNAiLDH-A100

  • RNAi vs. HIVRNAi

  • HIV reverse transcriptaseHIV siRNA-siRNA

  • Reference (siRNA Design Guidelines) (siRNA Technology) (siRNA) (RNAi) (Nature journal)

  • Reference (Post-transcriptional regulation of microRNA expression) (DNA Methylation Regulates MicroRNA Expression) (Nobel Prize2006) (C.elegans from wikipedia) (photo from wikipedia)

  • Reference (2006--) () (RNA)

  • Reference (Introduction of a Chimeric Chalcone Synthase Gene into Petunia Results in Reversible Co-Suppression of Homologous Genes in trans) (A Similarity Between Viral Defense and Gene Silencing in Plants )(ribozyme ) (miRNA)

  • Reference!92AA8BE887664BCD!330.entry

  • Reference () () (The NEW ENGLAND JOURNAL of MEDICINE)