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Harvard iGEM 2005 BioWire & BioSketch. Orr Ashenberg, Patrick Bradley, Connie Cheng, Chris Doucette, Hing Eng, Jennifer Gao, Kang-Xing Jin, Yin Li, Thomas Noriega, Danny Popper, Sasha Rush, Yves Wang 2005-08-24. The Projects. BioWire Propagating pulse generator BioSketch
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Harvard iGEM 2005BioWire & BioSketch Orr Ashenberg, Patrick Bradley, Connie Cheng, Chris Doucette, Hing Eng, Jennifer Gao, Kang-Xing Jin, Yin Li, Thomas Noriega, Danny Popper, Sasha Rush, Yves Wang 2005-08-24
The Projects • BioWire • Propagating pulse generator • BioSketch • Bacterial sketch pad
BioWireThe Wire Made of Biology Orr, Patrick, Connie, Kang-Xing, Danny, Sasha
BioSketchThe Bacterial Sketch Pad Chris, Hing, Jenny, Yin, Thomas, Yves
Goals • Make a sketch pad with bacteria • Write with a UV pen • Erase with heat • Advance synthetic biology • Rationally design & build genetic switches
Project Concept • Animation
Pen Eraser Eraser Circuit Design(s) • See also: Animation Pen Ink Protein A Protein B
Results (l CI-LacIts Switch) • UV turns switch ON • Heat turns switch OFF 48 J/m2 o.n. @ 40C
Problems with the Switch • ON state • Not complete: Many cells still OFF • "ON" cells are not fluorescent enough (in fact, not visible to naked eye) • State maintenance • States maintained at 4h • Begin to return to basal state by 1day later
Future Directions • l CI-LacIts switch • Get better induction with UV • Achieve more robust state-maintenance • Draw/erase on a lawn • 434 CI-l CIts switch • Assemble from BlueHeron-synthesized parts • Other reporters
Reflections • Planning & organization • Tracking assembly progress: Sasha's database for molecular cloning • Things take longer than expected • Shift focus from assembly to design/testing • Sequence as you go • The Registry • Incomplete part annotation • Experience issues of subparts should be annotated in superparts • Time of last update might be useful • Non-standard part requests • It's much better now than @ the beginning of the summer
Acknowledgments • Alain Viel, Ira Phillips, Sasha Wait • Pam Silver, George Church, Kit Parker, Radhika Nagpal • Harvard Provost Office, HHMI, DEAS, Division of Life Sciences • Parts Registry at MIT • Weiss Lab • Ron Weiss, Subhayu Basu, Allen Hsu, David Braun • Whitesides Lab • George Whitesides, Douglas Weibel, Andrew Lee, Scott Potenta, Mike Fuerstman • Parker Lab • Po-Ling ??, Josh ??, Daniel Goodman • CNS Facility • Yuan Lu, Steve Shepard • Collins Lab • Jim Collins, Hideki Kobayashi, Mads Kaern • Dana Farber Flow Cytometry Facility • Kat ?? • Bauer Center Flow Cytometry Facility • Brian Tilton