Timely E. coli Engineering a standardised cellular oscillator - - PowerPoint PPT Presentation

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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

SLIDE 1

Timely E. coli

Engineering a standardised cellular

scillator

UQ-Australia iGEM 2011 Regional Jamboree - Asia

Inspiration Circuitry The Future Modelling

SLIDE 2

Our Team

Inspiration Circuitry The Future Modelling

SLIDE 3

Our Team

Inspiration Circuitry The Future Modelling

SLIDE 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 The Future Modelling

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Effects of the circadian rhythm

Inspiration Circuitry The Future Modelling

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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

Hypertension.2005; 45: 874-879 Hartman et al. Am J Physiology. 1991; 260: E100-110

Inspiration Circuitry The Future Modelling

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Human circadian circuit

Gallego et al. Molecular Cell Biology. 2007; 8: 139-148

Key features:

Time delay
Negative feedback loop

Inspiration Circuitry The Future Modelling

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Mammalian Model

Tigges et al. Nature. 2009; 457: 309-312

Inspiration Circuitry The Future Modelling

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Prokaryotic Models

Atkinson et al. Cell. 2003; 113: 597-607

Inspiration Circuitry The Future Modelling

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Prokaryotic Models

Stricker et al. Nature. 2008; 456: 516-520

Simpler circuit
Dual response promoter

Inspiration Circuitry The Future Modelling

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Our system

araC glnG lacI

glnAP2 pBAD plac/ara

Inspiration Circuitry The Future Modelling

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glnG GlnG

S P

pSB1C3

Inspiration Circuitry The Future Modelling

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araC araC

S P

pSB1C3

Inspiration Circuitry The Future Modelling

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lacI lacI

S P

pSB1C3

Inspiration Circuitry The Future Modelling

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Mathematical Model

araC glnG lacI

glnAP2

pBAD plac/ara

Inspiration Circuitry The Future Modelling

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Model Analysis

Modelling of a novel

synthetic system

Kinetics sourced from pre-

existing work

Limited by differences

in experimental conditions

Inspiration Circuitry The Future Modelling

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Gene dosage effects

Tigges et al. Nature. 2009; 457: 309-312

Inspiration Circuitry The Future Modelling

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Chromosome integration

1. Landing pad
Use a pre-determined landing site to

target our constructs

2. Homologous recombination
Via identity with the landing pad site
3. Integration!
Now we have no need to worry about

fluctuations in gene dosage

Kuhlman et al. Nucleic Acids Research. 2010; 38(6): e92

1 1 3 2

Inspiration Circuitry The Future Modelling

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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 The Future Modelling

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…And along with the science

Biofutures + pain

Inspiration Circuitry The Future Modelling

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Share your human

practices and

utreach activities
Catch up (and

collaborate!) with iGEM’ers …And along with the science

Inspiration Circuitry The Future Modelling

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UQ iGEM

Oscillatory circuits are

important in nature

We are standardizing the

biological ‘clock’

Outreach is an important part
f science too!

Inspiration Circuitry The Future Modelling

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Thank You

Inspiration Circuitry The Future Modelling

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Kuhlman & Cox (2010) Chromosome Integration

Inspiration Circuitry The Future Modelling