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Team MembersUndergraduate 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)
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Two Designs
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Temperature (°C)
32
Bacterial Etch-A-SketchCell See Us Temperature Sensor
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Cell See Us Tuneable Temperature Sensor: Design
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R0010
Rfp
R0011
I12007
GfpTemperature
Sensitive Dimerization
LacICI
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R0010
Rfp
R0011
I12007
GfpTemperature
Sensitive Dimerization
LacICI
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R0010 R0011
I12007
Temperature Sensitive
Dimerization
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R0010 R0011
I12007
Temperature Sensitive
Dimerization
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R0010 R0011
I12007
Temperature Sensitive
Dimerization
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Cell See Us Tuneable Temperature Sensor
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Cell See Us Tuneable Temperature Sensor
T1
T2
T3
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Cell See Us Tuneable Temperature Sensor
T1
T2
T3
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Cell See Us Tuneable Temperature Sensor
T1
T2
T3
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Cell See Us Tuneable Temperature Sensor
T1
T2
T3
Transcription rate at T1
Transcription rate at T2
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34 36 38 40 42
Temperature (°C)
32
34 36 38 40 42
10000
20000
30000
40000
Temperature (°C)
Rfp
Gfp
nM
lacI fp1 fp2CI
Increase in temperature decreases the repression by lacI
Modeling Results
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Bacterial Etch-A-Sketch: Design
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Bacterial Etch-A-Sketch Write/Draw with Pipette filled with Lactose Design will glow green
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Bacterial Etch-A-Sketch Erase using pipettefilled with Tetracyclin Picture will cease
to glow green
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Bacterial Etch-A-Sketch
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R0011
Gfp
R0040
Lactose Binds with LacI
Inactive Repressor
TetRLacZ
(Metabolizes Lactose)LacI
Lactose
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R0011 R0040
TetRLacZ
(Metabolizes Lactose)
Lactose Binds with LacI
Inactive Repressor
Lactose
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R0011 R0040
TetRLacZ
(Metabolizes Lactose)
Tetracyclin Binds with
TetR
Inactive Repressor
aTc
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R0011 R0040
LacZ(Metabolizes Lactose)
Tetracyclin Binds with
TetR
Inactive Repressor
aTc
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R0011 R0040
LacZ(Metabolizes Lactose)
LacZ metabolizes
Lactose
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R0011 R0040
LacZ(Metabolizes Lactose)
LacZ metabolizes
Lactose
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R0011
Gfp
R0040
TetRLacZ
(Metabolizes Lactose)LacI
LacZ metabolizes
Lactose
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Project Plan and Execution
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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
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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
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Initial Assembly Schedule Part Week 1 Week 2 Week 3 Week 4
I0500
J06801
B0031
C0056
B0015
I12006
E0240
J04450
BBa_J11022
Testing
R0040
E0433
R0011
E0422
BBa_J11031
Testing Testing
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Parts Made
BBa_J11042 BBa_J11051 B0031+ C0056 R0040+ E0433
BBa_J11052R0011+ E0422
And possibly:
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Lessons for Next Year
• Larger team• Schedule undergraduate lab time more
effectively• Prepare team for assembly
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Questions?