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iGEM @ Imperial Starting off: Week 1 Week 1 Engineering/Biology Introduction Lectures Journal Club Wiki Brainstorming 3 ideas Bio Hard Drive Remote Controlled Bacteria Bio Calculator Bio-Clock Cell Networking Crazy Ideas Bacterial balloon Biological to Electrical Interface Only three ideas made the cut. Bio-Clock (Re-defining Time) • Pulse of AHL moves along a gutter of medium • Cells fluoresce when activated •Refractory period •The time period is controlled by •the radius •gutter width •cell density Bio-Memory •Data stored in bacteria written using a green laser •Data read using a UV laser + fluorimeter •Cells either 1 (RFP) or 0 (no RFP) •Data stored in switch Very High Compression due to small size of bacteria The Oscillator Culture Wide oscillations of AHL Frequency must be tuned easily Oscillations must be stable Changing Concentration of AHL Week 2 Investigation of all 3 ideas Modelling Evaluating risks Start work in the Wet lab Decision on the Oscillator as main project; can use other ideas as further developments Week 3 Further research Modelling Assembly of parts Protocols for testing parts Setting up OWW Predator-Prey Inte Predator Prey Dynamics Stabl Lotka-Voltarra Model output •Preda •The on species •The gr solely d •The P growth carryin •The P Design A A • Positive Feedback of A • AB Induces production of more B • Both A and B are used to make AB B B Two Cell System Two independent populations of Cells These cells do not kill each other A B Altering the initial ratios of these cells will alter the frequency of oscillations Design Cell1 (Prey) Prey cell must produce molecule A exponentially Lux R is produced which detects molecule A Pc Lux R Then initiates transcription at Plux Pc is always on A A Plux Lux I A A Which Produces More A Design The Predator Cell The role of a predator is to reduce the prey numbers as a function of the predator population numbers. Predator Detect Prey Population Size Reduce Prey Population Size Design The Predator Cell Detects Prey Population Size Plux Lux R Reduces Prey Population Size aiiA Design The Predator Cell LuxR A A LuxR Plux A Lux R aiiA aiiA Design (Entire System) P c A Lux R Diffusion A The Prey Cell Plu x Extra cellular pool of A (HSL) (this should oscillate) Lux I A The Predator Cell Lux R A A Lux R Plu x Lux R aii A aiiA Modelling Tom’s Monster Modelling Testing parts: T9002 A shocking discovery: at first sight... After finishing our oscillator design... MIT Project 2004: Cell-Cell synchronized Oscillator Design Similar approach using concepts of quorum sensing BUT: This system does not use predator-prey dynamics and is implemented in a single cell (ours is multicellular MIT Oscillator Design http://web.mit.edu/~cbatten/www/work/ ssbc04/system-spec-ssbc04.pdf Communication: The Wiki Wiki-Newspaper Documentation for future references Communication Within the team With other teams Monitoring progress (Gantt Chart) Present ourselves & our project Outline Further modelling & testing of parts Parts assembly Phase 2 Coupling the oscillator to a biological to electrical interface Synchronizing oscillations 2 petri dishes Thank you