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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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Presentation Transcript
team members
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)

two designs

32

34

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Temperature (°C)

Two Designs

Cell See Us

Temperature Sensor

Bacterial Etch-A-Sketch

slide5

Gfp

I12007

CI

LacI

Rfp

R0010

R0011

Temperature Sensitive Dimerization

slide6

Gfp

I12007

CI

LacI

Rfp

R0010

R0011

Temperature Sensitive Dimerization

slide7

R0010

R0011

Temperature Sensitive Dimerization

I12007

slide8

R0010

R0011

Temperature Sensitive Dimerization

I12007

slide9

R0010

R0011

Temperature Sensitive Dimerization

I12007

cell see us tuneable temperature sensor4

T1

T2

T3

Cell See Us Tuneable Temperature Sensor

Transcription rate at T1

Transcription rate at T2

slide15

fp2

fp1

lacI

CI

Rfp

40000

nM

30000

Gfp

20000

10000

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40

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36

38

40

42

Temperature (°C)

Temperature (°C)

Modeling Results

Increase in temperature decreases the repression by lacI

bacterial etch a sketch
Bacterial Etch-A-Sketch

Write/Draw with Pipette

filled with Lactose

Design will glow green

bacterial etch a sketch1
Bacterial Etch-A-Sketch

Erase using pipette

filled with Tetracyclin

Picture will cease

to glow green

slide20

LacZ

(Metabolizes Lactose)

LacI

Gfp

TetR

R0011

R0040

Lactose Binds with LacI

Lactose

Inactive Repressor

slide21

LacZ

(Metabolizes Lactose)

TetR

R0011

R0040

Lactose Binds with LacI

Lactose

Inactive Repressor

slide22

LacZ

(Metabolizes Lactose)

TetR

R0011

R0040

Tetracyclin Binds with TetR

aTc

Inactive Repressor

slide23

LacZ

(Metabolizes Lactose)

R0011

R0040

Tetracyclin Binds with TetR

aTc

Inactive Repressor

slide24

LacZ

(Metabolizes Lactose)

R0011

R0040

LacZ metabolizes Lactose

slide25

LacZ

(Metabolizes Lactose)

R0011

R0040

LacZ metabolizes Lactose

slide26

LacZ

(Metabolizes Lactose)

LacI

Gfp

TetR

R0011

R0040

LacZ metabolizes Lactose

initial plan
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
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
Parts Made

BBa_J11042 BBa_J11051

B0031+ C0056R0040+ E0433

And possibly:

BBa_J11052

R0011+ E0422

lessons for next year
Lessons for Next Year
  • Larger team
  • Schedule undergraduate lab time more effectively
  • Prepare team for assembly