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( + ). Team Members. Undergraduate Students Aman Arya , Biology (Waterloo) Joseph Chan , Molecular Genetics (Toronto) Hannah Fong , Immunology / Chemistry (Toronto) Lutz Kummer , visiting Biology (Waterloo) Emanuel Nazareth , Biomedical Engineering (Toronto)
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Team Members Undergraduate Students Aman Arya, Biology (Waterloo) Joseph Chan, Molecular Genetics (Toronto) Hannah Fong, Immunology / Chemistry (Toronto) Lutz Kummer, visiting Biology (Waterloo) Emanuel Nazareth, Biomedical Engineering (Toronto) Martina Spranger, visiting Biology (Waterloo) Max Wong, Biomedical Engineering (Toronto) Graduate Students Seema Nagaraj, IBBME and Dept. of Electrical Engineering (Toronto) Matt Scott, Dept. of Applied Mathematics (Waterloo) Cindy Ying Yin Yip, IBBME (Toronto) Technical Support Andrew Hessel, Lab Manager, Mak Lab, University Health Network Faculty Advisors Prof. Stephen Davies, IBBME and Dept. of Electrical Engineering (Toronto) Prof. Brian Ingalls, Dept. of Applied Mathematics (Waterloo)
32 34 36 38 40 42 Temperature (°C) Two Designs Cell See Us Temperature Sensor Bacterial Etch-A-Sketch
Gfp I12007 CI LacI Rfp R0010 R0011 Temperature Sensitive Dimerization
Gfp I12007 CI LacI Rfp R0010 R0011 Temperature Sensitive Dimerization
R0010 R0011 Temperature Sensitive Dimerization I12007
R0010 R0011 Temperature Sensitive Dimerization I12007
R0010 R0011 Temperature Sensitive Dimerization I12007
T1 T2 T3 Cell See Us Tuneable Temperature Sensor
T1 T2 T3 Cell See Us Tuneable Temperature Sensor
T1 T2 T3 Cell See Us Tuneable Temperature Sensor
T1 T2 T3 Cell See Us Tuneable Temperature Sensor Transcription rate at T1 Transcription rate at T2
fp2 fp1 lacI CI Rfp 40000 nM 30000 Gfp 20000 10000 34 36 38 40 42 32 34 36 38 40 42 Temperature (°C) Temperature (°C) Modeling Results Increase in temperature decreases the repression by lacI
Bacterial Etch-A-Sketch Write/Draw with Pipette filled with Lactose Design will glow green
Bacterial Etch-A-Sketch Erase using pipette filled with Tetracyclin Picture will cease to glow green
LacZ (Metabolizes Lactose) LacI Gfp TetR R0011 R0040 Lactose Binds with LacI Lactose Inactive Repressor
LacZ (Metabolizes Lactose) TetR R0011 R0040 Lactose Binds with LacI Lactose Inactive Repressor
LacZ (Metabolizes Lactose) TetR R0011 R0040 Tetracyclin Binds with TetR aTc Inactive Repressor
LacZ (Metabolizes Lactose) R0011 R0040 Tetracyclin Binds with TetR aTc Inactive Repressor
LacZ (Metabolizes Lactose) R0011 R0040 LacZ metabolizes Lactose
LacZ (Metabolizes Lactose) R0011 R0040 LacZ metabolizes Lactose
LacZ (Metabolizes Lactose) LacI Gfp TetR R0011 R0040 LacZ metabolizes Lactose
Initial Plan • May – June • Brainstorm and finalize design • July – August • Obtain constructs and test • September – October • Make revisions, fine tune model parameters, obtain and test new constructs
Actual Execution • May – August • Brainstorm designs • September • Found out we needed to prepare constructs ourselves • Reconfigure designs to use available parts • Recruitment of volunteers with wet lab experience • October – Early Nov. • Undergraduates began assembly • Coincided with peak of undergrad midterm season
Parts Made BBa_J11042 BBa_J11051 B0031+ C0056R0040+ E0433 And possibly: BBa_J11052 R0011+ E0422
Lessons for Next Year • Larger team • Schedule undergraduate lab time more effectively • Prepare team for assembly