Nobel Prize 2006

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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

    mRNARdRpdsRNAsiRNARNAi

  • 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

  • Referencehttp://www.ambion.com/techlib/tb/tb_506.html (siRNA Design Guidelines) http://www.genscript.com/siRNA_technology.html (siRNA Technology)http://www.ambion.com/techlib/tn/95/952.html (siRNA)http://content.nejm.org/cgi/content/full/347/17/1364 (RNAi)http://www.nature.com/nature/journal/v391/n6669/pdf/391806a0.pdf (Nature journal)

  • Referencehttp://rnajournal.cshlp.org/content/12/7/1161.full.pdf (Post-transcriptional regulation of microRNA expression)http://www.landesbioscience.com/journals/cbt/article/HanCBT6-8.pdf (DNA Methylation Regulates MicroRNA Expression)http://nobelprize.org/nobel_prizes/medicine/laureates/2006/index.html (Nobel Prize2006)http://en.wikipedia.org/wiki/C_elegans (C.elegans from wikipedia)http://upload.wikimedia.org/wikipedia/commons/b/be/CrawlingCelegans.gif (photo from wikipedia)

  • Referencehttp://nobelprize.org/nobel_prizes/medicine/laureates/2006/fire-autobio.html()http://www.biox.cn/content/200610/20061002180126_322511.shtml (2006--)http://nobelprize.org/nobel_prizes/medicine/laureates/2006/mello-autobio.html()http://web1.nsc.gov.tw/ct.aspx?xItem=8539&ctNode=40&mp=1 ()http://www.newsoo.cn/epaper/yyjjb/2007/05/18/484274.htm (RNA)

  • Referencehttp://www.plantcell.org/cgi/reprint/2/4/279.pdf (Introduction of a Chimeric Chalcone Synthase Gene into Petunia Results in Reversible Co-Suppression of Homologous Genes in trans)http://www.sciencemag.org/cgi/content/full/276/5318/1558 (A Similarity Between Viral Defense and Gene Silencing in Plantshttp://porpax.bio.miami.edu/~cmallery/255/255life/ribozyme.jpg )(ribozyme )http://www.genscript.com/images/sirna/miRNA.jpg (miRNA)

  • Reference http://en.wikipedia.org/wiki/Dicer http://en.wikipedia.org/wiki/RNA_interference http://en.wikipedia.org/wiki/RNA-induced_silencing_complex http://huiweno824.spaces.live.com/blog/cns!92AA8BE887664BCD!330.entry http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=15845026 http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1370384

  • Referencehttp://www.afv.org.hk/AMD_c.htm ()http://www.ebiotrade.com/newsf/2008-8/200888154931.htm ()http://content.nejm.org/cgi/content/full/347/17/1364/F2 (The NEW ENGLAND JOURNAL of MEDICINE)http://www.medscape.com/pi/editorial/cmecircle/2000/117/baldassano1/slide27.gif

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