Team
We are a team of seven highly motivated Bachelor- and Master Students at the ETH Zurich pursuing various fields such as Biotechnology, Biomedical Engineering, Neurobiology and Bioinformatics. The iGEM project is carried out at one of the youngest departments of ETHZ located in Basel-Department of Biosystems Science and Engineering - flourishing in interdisciplinary biological research. If you're around Basel, make sure to visit our team's lab to play the bio-game Colisweeper!

From Minesweeper to Colisweeper
Our project-Colisweeper- is a biological version of the computer game Minesweeper. The minefield is replaced by mine and non-mine colonies on an agar plate. Quorum sensing molecule OHHL communicates between colonies which are designed to express different orthogonal hydrolases depending on the signal processed. By pipetting a colorless multi-substrate mix onto mine or non-mine colonies, color appears which indicates the next logical move for the player. If you are certain of a mine colony, you can use a second substrate solution to flag the mine.

Information Processing
Our non-mine colonies are designed to distinguish between different concentrations of OHHL that translate this information into expression of different sets of hydrolases. They are equipped with mutated LuxR promoters with different OHHL sensitivities which serve as highpass filters. The promoters were created using site-saturation mutagenesis. Through mutation of the LuxR binding sites we were able to tune the promoters to different OHHL affinities .

Hydrolase Reactions
We use a set of orthogonal hydrolases as our reporter system that react within minutes with the added multi-substrate to produce a visible color. The set of hydrolases such as alkaline phosphatase (phoA), β-galactosidase (lacZ), acetylesterase (aes), β-N-Acetylglucosaminidase (nagZ) and β-glucuronidase (gusA) and their respective substrates react to achieve fast and colorful outputs with each color indicative of the next logical move for the player.

The Model
As our system is based on quorum sensing of OHHL, the diffusion of OHHL in the mine field is a vital part of the system. We study the diffusion in our system in a hexagonal grid format with mine and non-mine colonies using a spatio-temporal model. The information from the model was used to validate and improve our system.

Experiments
Starting with characterization of the OHHL diffusion in agar plates we went on to test different sender- and receiver-cell pairs in the hexagonal grid. Initially we used Green fluorescent Protein (GFP) as a reporter to study the OHHL interaction between mine and non mine colonies. Together with promoter mutagenesis and screening, we also tested and characterized different hydrolase-substrate pairs which are crucial to get the game going!

Human practices
As an inspiration from our project, we analyzed the relationship between synthetic biology and games. For one thing synthetic biology can be used to play common games in a new way, possibly for educational purposes or as a basis for proof-of-principle experiments for new circuits. More recently synthetic biologists also started to use games as a research tool, an innovative approach to make use of crowd-sourcing and distributed computing. We want to find correlations and discuss possible consequences for Synthetic Biology.