(+)(+). Team Members Undergraduate Students Aman Arya, Biology (Waterloo) Joseph Chan, Molecular...

( + )

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)

Two Designs

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Temperature (°C)

32

Bacterial Etch-A-SketchCell See Us Temperature Sensor

Cell See Us Tuneable Temperature Sensor: Design

R0010

Rfp

R0011

I12007

GfpTemperature

Sensitive Dimerization

LacICI

R0010

Rfp

R0011

I12007

GfpTemperature

Sensitive Dimerization

LacICI

R0010 R0011

I12007

Temperature Sensitive

Dimerization

R0010 R0011

I12007

Temperature Sensitive

Dimerization

R0010 R0011

I12007

Temperature Sensitive

Dimerization

Cell See Us Tuneable Temperature Sensor

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

T1

T2

T3

Transcription rate at T1

Transcription rate at T2

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

Bacterial Etch-A-Sketch: Design

Bacterial Etch-A-Sketch Write/Draw with Pipette filled with Lactose Design will glow green

Bacterial Etch-A-Sketch Erase using pipettefilled with Tetracyclin Picture will cease

to glow green

Bacterial Etch-A-Sketch

R0011

Gfp

R0040

Lactose Binds with LacI

Inactive Repressor

TetRLacZ

(Metabolizes Lactose)LacI

Lactose

R0011 R0040

TetRLacZ

(Metabolizes Lactose)

Lactose Binds with LacI

Inactive Repressor

Lactose

R0011 R0040

TetRLacZ

(Metabolizes Lactose)

Tetracyclin Binds with

TetR

Inactive Repressor

aTc

R0011 R0040

LacZ(Metabolizes Lactose)

Tetracyclin Binds with

TetR

Inactive Repressor

aTc

R0011 R0040

LacZ(Metabolizes Lactose)

LacZ metabolizes

Lactose

R0011 R0040

LacZ(Metabolizes Lactose)

LacZ metabolizes

Lactose

R0011

Gfp

R0040

TetRLacZ

(Metabolizes Lactose)LacI

LacZ metabolizes

Lactose

Project Plan and Execution

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

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

Parts Made

BBa_J11042 BBa_J11051 B0031+ C0056 R0040+ E0433

BBa_J11052R0011+ E0422

And possibly:

Lessons for Next Year

• Larger team• Schedule undergraduate lab time more

effectively• Prepare team for assembly

Questions?