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 gene RNA Polymerase DNA Promoter

8. Gene Expression gene RNA Polymerase DNA mRNA Promoter

9. Gene Expression gene RNA Polymerase Ribosome DNA mRNA Promoter

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

11. Gene Networks

12. transcription factor (repressor) X

13. Gene Networks transcription factor (activator)

14. Genetic Amplifier Topology cI cro gfp P lac P tet P RM - (+) activator (-) repressor

15. Wild Type Promoter gfp P RM

16. Double Mutation gfp P RM

17. Implementation InputAmplifier

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

22. Experimental Results High Copy Number

23. Experimental Results Low Copy Number

24. Conclusions Simulation: low copy number amplifier based on double- mutant P RM 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