Sources and Inspiration

Our circuit is largely based off the following papers and groups:

the work with toehold switches, cell-free systems, and paper-based sensors detailed in "Paper-Based Synthetic Gene Networks" by Pardee et al., which we consistently refer to as "the Collins paper";

the 2015 Pitt iGEM team and their work with cell-free, paper-based systems and the Collins materials;

the thallium DNAzyme described by Huang, Vazin, and Liu in "Desulfurization Activated Phosphorothioate DNAzyme for the Detection of Thallium";

the amplification systems developed by Wang, Barahona, and Buck in "Engineering modular and tunable genetic amplifiers for scaling transcriptional signals in cascaded gene networks"; and

toehold switch design described by the 2015 Exeter iGEM team.

Labs

Huge thank-you's go to the following groups and people for making this year's iGEM team a success:

Dave Gau from the Roy lab, who provided us with the various odds and ends we needed to grow cells;

Steve from the Shroff lab, who provided us with reagents and materials for making buffers, a freezer, and friendliness every day;

Neill Turner from the McGowan Institute for giving us access to a plate reader; and

Nick Ankenbruck from the Deiters lab, who helped us run dPAGE gels.

Advisors and Mentors

Our advisors and mentors met with us every week to discuss ideas for the circuit, experimental results, experimental design, and presentations. Special thanks go to:

Dr. Jason Lohmueller, who started iGEM at Pitt and provided valuable insights for experimental designs and interpretation of results;

Dr. Sanjeev Shroff, who worked out all the logistics to make iGEM possible;

Dr. Natasa Miskov-Romanov, who provided us with a lab space;

Dr. Alex Deiters, who always knew we could do better; and, last but not least,

Dr. Lisa Antoszewski. Without her dedication, patience, and organization as our laboratory mentor, we would never have passed the theoretical stages.

Team

Sponsors

Thank you to Breton Hornblower and Cory Tuckey from NEB for making PURExpress a viable option for our experiments.

Project Timeline

The team met for the first time at the beginning of April. For the next month-and-a-half, we brainstormed ideas for our project. By our first day in lab at the end of May, we knew we wanted to design a thallium sensor. Our circuit design solified over the next week or so as we started to familiarize ourselves with the lab space. By the next week, we were learning lab techniques and applying them to our project. See our lab notebook for more details on what we worked on each week.