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internationalregulomeconsortium

www.internationalregulomeconsortium.ca. 1. Capable of long term self-renewal. 2. Capable of multipotent differentiation. BDNF FGF Shh Wnts BMPs. Oligodendrocyte. Neuron. Astrocyte. Neural Stem Cell. Definition of a Stem Cell. Ectoderm. Mesoderm. Zygote. Blastocyst.

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internationalregulomeconsortium

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  1. www.internationalregulomeconsortium.ca

  2. 1. Capable of long term self-renewal 2. Capable of multipotent differentiation BDNF FGF Shh Wnts BMPs Oligodendrocyte Neuron Astrocyte Neural Stem Cell Definition of a Stem Cell

  3. Ectoderm Mesoderm Zygote Blastocyst Endoderm PGC The Transcriptome Problem

  4. IRC Background • August 2003, First Discussions on Project • Stuart McKeen (Government of Ontario) • Ronnie Gavsie (Ontario Genomics Institute) • Aled Edwards (Structural Genomics Consortium) • Seek international partners to form consortium • France, UK, Singapore • Steering Committee Workshop, Ottawa 15/12/2003 Michael Rudnicki, Jack Greenblatt, Irwin Davidson, Bing Lim, Janet Rossant, Lazslo Tora, Pierre-Marie Lledo, Jacques Hatzfeld, Miguel Andrade, Lynn Megeney, • Additional Partners Sought • European Union • Australia • Founding Workshop of IRC, Ottawa 3/5/2004 • Australia 75 participants from 8 countries

  5. Dr. Michael A. Rudnicki Ottawa Health Research Institute, Canada Dr. Irwin Davidson IGBMC, France Dr. William Skarnes Wellcome Trust Sanger Institute, U.K. Dr. Ihor Lemischka Princeton University, USA Dr. Jack Greenblatt C.H. Best Institute, Toronto, Canada Dr. Frank Grosveld Erasmus University, The Netherlands Dr. Bing Lim Genome Institute of Singapore Dr. John Mattick University of Queensland, Australia Dr. Janet Rossant Samuel Lunenfeld Research Institute, Toronto, Canada Steering Committee

  6. The Vision The Vision A complete understanding of the transcriptional nodes and networks that regulate all mammalian cell biology A complete understanding of the transcriptional nodes and networks that regulate all mammalian cell function

  7. The Rational • IRC working group has found that the mouse genome contains about 2,500 identifiable transcription factors • Combinatorial interactions among these proteins and the activation or repression of gene transcription forms a complex of poorly understood regulatory nodes and networks • The cascades of regulatory interactions between transcription factors together define all aspects of mammalian biology

  8. Scientific Objectives • Undertake the complete genetic description of stem cell function, lineage commitment, and differentiation • Characterize the protein components of transcriptional complexes containing all potential transcriptions factors • Identify and validate the complete set of transcription factor binding sites and corresponding target genes • Derive computational models that describe the hierarchal regulation of gene transcription

  9. Scientific Approach

  10. High Throughput Gene Targeting • Recombinational vector generation in BACs • TAP-tag and flox up to 2000 genes in mice • ES/embryo aggregation

  11. Wild-type Oct4 Allele 5.16 kb Targeted Oct4 Allele Flag-TEV-His Neo Targeted Oct4 Allele (after FLP Recombinase) Flag-TEV-His FRT Site

  12. Mass Spectroscopy • Tandem affinity purification of complexes • LC MS MS Transcription Complex

  13. A – Build construct C– Purify complex Associated proteins C-terminal TAP tag N C Ni+ resin a-FLAG M2 6xHis TEV 3xFLAG TEV Protease B – Express construct in cells Elute Pax7 Pax7 Pax7 Pax7 TAP Tag Methodology

  14. Identification of Target Genes • Chromatin Immunoprecipitation (ChIP) by Tandem Affinity Purification • Di-Tag DNA sequencing of ChIP’d DNA

  15. Computational Analysis • Generation of models of regulation of transcriptome states Six1 Mef2C/D Myf5/MyoD Myogenin Muscle Genes Pax7 Stem Cell Muscle Progenitor New Muscle fiber • Pax7 • Six1 • Myf5 • MyoD • Myogenin • Mef2C • Mef2D

  16. IRC Current Status • Founding of the International Regulome Consortium at May 2004 Workshop in Ottawa • Delivered Ottawa White Paper outlining the objectives and methodology of the IRC July 2004 • Non-profit corporation for governance and management May 2005 • Experimental work underway in participating Institutions • Seed funding awarded in Canada (CIHR/MEDT) and France (MR) in 2004/2005 • Sanger Center initates Regulome with $8.8M/4years September 1, 2005 • Full applications pending in Canada (ICI, CFI), France and EU 6th Framework • Incoporation as Not-for-Profit November 2005 • Official launch in March 2006

  17. IRC Integrated Work Plan

  18. Management Structure

  19. Summary • Third generation genomics project that will revolutionize our understanding of cell function at the genomic level • Canadian-led, multinational scientific consortium - 17 institutions in 6 countries • Facilitate development of novel therapeutics for regenerative medicine • Profound human health, socio-economic implications www.internationalregulomeconsortium.ca

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