1. iGEM 2005: Team BioWire and BioLoserz!!! LOL Orr Ashenberg, Patrick Bradley, Connie Cheng, Kang-Xing Jin, Danny Popper, Sasha Rush

2. Project Overview Goal: to engineer a synthetic, macroscopic biological “wire,” capable of propagating a chemical pulse down its length – Chemical signaling uses homoserine lactones (quorum sensing pathway in V. fischeri) – Pulse is controlled via transcriptional regulation – Bacteria shaped into wire using photolithography

3. BioWire: “Pulse” Propagation Initial Signal

4. Circuit Design: Pulse Generator Incoherent feed-forward loop – AHL stimulates production of both cI and GFP – cI represses GFP – Result: a pulse of fluorescence, followed by a refractory period cI GFP AHL

5. Circuit Design: BioWire Incoherent feed-forward loop plus positive feedback – LuxI produces more AHL, which triggers neighboring cells cIGFP & LuxIAHL

6. Timing Diagram for Pulse YFP cI AHL

7. Successful Experiments

8. Future Work

9. Parts Constructed

10. Sender / Receiver Phase Sender / Receiver GFP Receiver Phase Receiver GFP

11. Senders, 100X phase Senders, 100X GFP Receivers, 100X phase Receivers, 100X GFP

12. Receivers near senders, 100X phase Receivers near senders, 100X GFP Receivers far from senders, 100X phase Receivers far from senders, 100X GFP