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A Genetic Differential Amplifier: Design, Simulation, Construction and Testing

A Genetic Differential Amplifier: Design, Simulation, Construction and Testing

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A Genetic Differential Amplifier: Design, Simulation, Construction and Testing

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  1. A Genetic Differential Amplifier: Design, Simulation, Construction and Testing Seema Nagaraj and Stephen Davies University of Toronto Edward S. Rogers Sr. Department of Electrical and Computer Engineering Institute of Biomaterials and Biomedical Engineering

  2. Engineered Genetic Circuits • artificial networks of interacting DNA, mRNA and proteins • behavior is analogous to electrical devices

  3. Genetic Differential Amplifier • differential amplifiers are fundamental components in many electronic circuits • a genetic differential amplifier could be a building block for more complex genetic circuits

  4. Electronic Differential Amplifier

  5. Amplifier Behaviour

  6. Gene Expression gene DNA Promoter

  7. Gene Expression RNAPolymerase gene DNA Promoter

  8. Gene Expression RNA Polymerase gene DNA Promoter mRNA

  9. Gene Expression RNA Polymerase gene DNA Promoter Ribosome mRNA

  10. Gene Expression RNA Polymerase gene DNA Promoter Ribosome mRNA Protein

  11. Gene Networks

  12. X Gene Networks transcription factor (repressor)

  13. Gene Networks transcription factor (activator)

  14. Genetic Amplifier Topology (+) activator cI Ptet gfp PRM - (-) repressor cro Plac

  15. gfp PRM Wild Type Promoter

  16. gfp PRM Double Mutation

  17. Implementation Input Amplifier

  18. Stochastic Modeling • includes inputs and output • system behaviour is described through reaction probabilities • system behaviour is simulated using the Gillespie algorithm

  19. Simulation Results(Low Copy Number)

  20. Simulation Results(High Copy Number)

  21. Experimental Results • Construct HIGH and LOW copy number variants of the circuit • Compare performance to simulated results

  22. Experimental ResultsHigh Copy Number

  23. Experimental ResultsLow Copy Number

  24. Conclusions Simulation: • low copy number amplifier based on double-mutant PRM yields “ideal” behaviour Experimental: • high copy number amplifier displays differential amplifier behaviour • low copy number amplifier displays positive results

  25. Acknowledgements • Anja Lowrance • Members of Davies Lab • Rogers Scholarship