Overview

On this page we explain the design principles we defined for our project and how we followed them. We structured our work in phases and tried to proceed through them. The phases apply to theoretical work (models) as well as to the practical (experiments). In the early phases, we learned to handle the subjects and get familiar with the theory and literature. We designed, ordered and built constructs for tests of the experimental procedure and for further optimization. In later phases, we tested predictions of the models and delivered parameters for new models. We engineered single parts to behave as our circuit requires by implementing predictions of the models. We combined functions if possible and showed that we can build more complex circuits with them.

We designed the project in a hierarchical bottom-up engineering approach: We divided the circuit into its different functions (F_a-F_e) and engineered them until they met our criteria:

• Tumor Sensor
• MRI Contrast Agent
• Anti-Cancer Toxin
• Heat Sensor
• Cell Lysis

The individual constructs were assembled with molecular cloning and the functions were tested with reporter genes such as gfp and mcherry. Only if they behaved according to our requirements, we combined functions together. In parallel we ordered as soon as possible the full genetic circuit of CATE with restriction sites along the critical loci in order to rapidly exchange promotors ribosome binding sites or coding sequences after we experimentally optimized the parts.