University of Toronto University of Waterloo CellsSeeUs Thermometer

University of Toronto & University of Waterloo Cells-See-Us Thermometer

Presentation Overview: • • • Project Overview Project Design and Philosophy Reaction Mechanisms Construction Results

Project Overview Cell-See-Us Thermometer: • a bio-synthetic device that displays changes in temperature through colour • red when hot, and green when cold Applications of a bio-synthetic thermometer • map temperature gradients with spatial resolution down to micrometers

Project Principle Temperature (T) Sensor Inverter Low Temperature: Green (GFP) High Temperature: Red (RFP) GFP: Green Flourescent Protein RFP: Red Flourescent Protein

Design Philosophy • Allow for modular testing of each vital component of the construct • Many external control points for system modulation • Construction flexibility

Reaction Mechanism Cold Gfp is formed Lac. I forms dimer Lac. I binds to Rfp-repressing site (R 0011) and tet. R-repressing site (R 0011). No Rfp or tet. R is made Hot Dimers break Lac. I monomers do not bind onto repressing sites – Rfp and tet. R are formed. tet. R inhibits Gfp

Deterministic Model:

Project Model - Concentration difference in reporter proteins between 27 o. C and 37 o. C

Construction Phase • Constructed 15 new parts (10 not shown)

Testing Phase Module 1 Module 2 Module 3 Cells-See-Us Thermometer

Module 1 – Reporter Functionality

Module 1 – Production of Lac. I

Module 1 – Temp. Dependence

Testing Phase Module 1 Module 2 Module 3 Cells-See-Us Thermometer

Module 2 – tet. R Functionality

Testing Phase Module 1 Module 2 Module 3 Cells-See-Us Thermometer

Module 3 – Temperature Test

Heat Gradient Plate Test

Before… And After…

Conclusions • Completed construction of designed genetic device • Basic functionality of the Cell-See-Us Thermometer was achieved: o Both red and green fluorescence o Temperature-dependent fluorescence levels

Acknowledgements Team Members: Sponsors: university of toronto engineeringsociety University of Waterloo Mathematics Endowment Fund Waterloo Engineering Endowment Fund Special thanks to: - University of Toronto Davies’ Lab for their generous donations of lab space, equipment, and supplies - Brian Ingalls for his generous support in modeling Thank you i. GEM organizers and MIT for hosting! Leaders: - Farshid Mirrahimi (Toronto) - Andy Hung (Toronto) - Charles Yoon (Toronto) - Nancy Xu (Waterloo) Active Members: - Niladri Chattopadhyay (Toronto) - Mohammed Hasan (Toronto) - Hari Vijayendran (Toronto) - Saba Mortazavi (Toronto) - Sara M Khojasteh (Toronto) Major Contributors and Organizers: - Vahe Akbarian (Toronto) - Melinda Yeung (Toronto) - Dave Puri (Toronto) - Wing. Yee Cheung (Toronto) - Irena Mahdavi (Toronto) - Natalie Yeung (Toronto) - Michael Leung (Toronto) - Konstantin Savitsky (York) - Christina Lucey (Waterloo) - Anne Tran (Waterloo) - Linda Chen (Waterloo) - Conrad Lochovsky (Toronto) - Rohan Gidvani (Waterloo) - Jovan Lukovich (Toronto) - Jeffrey Wong (Waterloo) - Nick Ngai (Toronto) Faculty Advisors: - Tara khiabani (Toronto) - Prof. Stephen Davies (Toronto) - Ho. Kwon Kim (Toronto) - Prof. Brian Ingalls (Waterloo) - Cheng. Chuan Qu (Toronto) - Seema Nagaraj (Toronto) - Adnan Najmi (Toronto) - Lance Da. Silva (Waterloo) - Elliott Sales de Andrade (Toronto) - Patrick Tsui (Waterloo) - Ting Zhu (Toronto) - Jessica Yang (Toronto) - George Ye (Toronto) - Ram Puri (Toronto) - Herak Sikder (Toronto) - Arash Mirrahimi (Toronto) - Sam Leung (Waterloo) - Chris Herriot (Waterloo)