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  • The Team
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• Our Project
  • Results Summary
  • Selection System
  • In-Vitro Characterization
  • In-Vivo Characterization
  • Microfluidics
  • Data
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• Tools
  • Gibson Assembly
  • MITOMI
• Notebook
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  • May
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• Considerations
  • Human Practices
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• Acknowledgements
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Welcome

It's on! After some warmup experiments during the semester, our team is now fully operational, and already having trouble getting away from the lab. Follow our adventures on twitter: IGEM_EPFL

Team, remember the Todo List

Project summary

Our project this summer was to set up a high-throughput method to create and characterize new transcription factors (TFs) that would recognize different promoter sequences. We mutated the TetR transcription factor and characterized in vitro their affinity to the consensus sequence (the Ptet promoter) as well as their position-weight matrices. The next step of the characterization is an in vivo readout system; we created and tested 2 different readout systems based on RFP expression, with either a positive or negative selection of the TetR-Ptet interaction. Wanting a high-throughput method, we decided also to use a lysis cassette as a reporter gene. The idea is to kill the cells in which a TetR mutant recognizes a Ptet sequence (either the consensus or a mutated one) and to recover their DNA; this can be coupled to microfluidics chemostat chambers, where hundreds of cell colonies can be grown at the same time.