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Timely E. coli Engineering a standardised cellular oscillator UQ-Australia iGEM 2011 Regional Jamboree - Asia Inspiration Circuitry Modelling The Future Our Team Inspiration Circuitry Modelling The Future Our Team Inspiration

  1. Timely E. coli Engineering a standardised cellular oscillator UQ-Australia iGEM 2011 Regional Jamboree - Asia Inspiration Circuitry Modelling The Future

  2. Our Team Inspiration Circuitry Modelling The Future

  3. Our Team Inspiration Circuitry Modelling The Future

  4. Circadian Rhythm Circadian rhythm regulates:  Sleep-wake cycle  Body temperature  Feeding behaviour and appetite  Hormone secretion and metabolism  Glucose homeostasis  Cell cycle progression Inspiration Circuitry Modelling The Future

  5. Effects of the circadian rhythm Inspiration Circuitry Modelling The Future

  6. Other effects 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Hartman et al. Am J Physiology. 1991; 260: E100-110 Hypertension.2005; 45: 874-879 Inspiration Circuitry Modelling The Future

  7. Human circadian circuit Key features: • Time delay • Negative feedback loop Gallego et al. Molecular Cell Biology. 2007; 8: 139-148 Inspiration Circuitry Modelling The Future

  8. Mammalian Model Tigges et al. Nature. 2009; 457: 309-312 Inspiration Circuitry Modelling The Future

  9. Prokaryotic Models Atkinson et al. Cell. 2003; 113: 597-607 Inspiration Circuitry Modelling The Future

  10. Prokaryotic Models • Simpler circuit • Dual response promoter Stricker et al. Nature. 2008; 456: 516-520 Inspiration Circuitry Modelling The Future

  11. Our system araC plac/ara glnG pBAD lacI glnAP2 Inspiration Circuitry Modelling The Future

  12. glnG GlnG P S pSB1C3 Inspiration Circuitry Modelling The Future

  13. araC pSB1C3 araC P S Inspiration Circuitry Modelling The Future

  14. lacI lacI P S pSB1C3 Inspiration Circuitry Modelling The Future

  15. Mathematical Model araC plac/ara glnG pBAD lacI glnAP2 Inspiration Circuitry Modelling The Future

  16. Model Analysis • Modelling of a novel synthetic system • Kinetics sourced from pre- existing work  Limited by differences in experimental conditions Inspiration Circuitry Modelling The Future

  17. Gene dosage effects Tigges et al. Nature. 2009; 457: 309-312 Inspiration Circuitry Modelling The Future

  18. Chromosome integration 1 1 1. Landing pad - Use a pre-determined landing site to target our constructs 2 2. Homologous recombination - Via identity with the landing pad site 3 3. Integration! - Now we have no need to worry about fluctuations in gene dosage Kuhlman et al. Nucleic Acids Research. 2010; 38(6): e92 Inspiration Circuitry Modelling The Future

  19. Applications • Standardised platform for modeling oscillating systems • Periodic medicine: potential drug or hormone delivery system • Biological Battery Cell: Generating current through periodic stimulation of ion channels Inspiration Circuitry Modelling The Future

  20. …And along with the science Biofutures + pain Inspiration Circuitry Modelling The Future

  21. …And along with the science • Share your human • Catch up (and practices and collaborate!) with iGEM’ers outreach activities Inspiration Circuitry Modelling The Future

  22. UQ iGEM • Oscillatory circuits are important in nature • We are standardizing the biological ‘clock’ • Outreach is an important part of science too! Inspiration Circuitry Modelling The Future

  23. Thank You Inspiration Circuitry Modelling The Future

  24. Kuhlman & Cox (2010) Chromosome Integration Inspiration Circuitry Modelling The Future

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