Module 1 builds synthetic haploid strains with refactored mating loci that are conjugated to make a functional IOD. These strains have the wild-type mating phenotype and differentially express a reprogrammed signalling pathway in their diploid state proving the feasibility of the clone-free assembly concept.

✓ Construct a set of reporter promoters for yeast cells

✓ Characterize reporter promoters in all mating types

✓ Design and materialized synthetic MATa and MATx strains

✓ Build a synthetic diploid strain with a functional yeast pheromone pathway

✓ Demonstrate the correct functionality of yeast pheromone pathway in synthetic diploids

Module 2 builds a set of orthogonal pheromones and receptors. These pheromone-receptor pairs enable specific localized signalling proving the feasibility of multichannel signal transmission underlying logic operations necessary for reliable diagnosis.

✓ Construct a set of yeast plasmids with different mating pheromones and their receptors and contribute to the Registry with BioBricks

✓ Verify the correct coupling of the receptors to the yeast pheromone mating pathway

✓ Verify the correct expression and secretion of the different pheromones

✓ Show the orthogonality of the used receptors and pheromones

Module 3 builds a set of location tags that recognize common tumor surface markers and agglutinate cell populations. Location tags displayed in the correct conformation strengthen cell-cell interactions to enable localization of signal transmission.

✓ Express streptavidin, EpCAM, Anti-EpCAM scFv, c-Myc scFv and anti-HuA scFv on the surface of yeasts

✓ Demonstrate the ability of our receptors to bind chosen markers

✓ Monitor the dynamic binding of our receptors and their corresponding markers

Module 4 provides modeling and simulation support for other modules.

✓ Develop a simulation environment CeCe to capture the complexity of cell-cell signal transmission

✓ Design an IOD chemical reaction network model

✓ Develop a schematic architecture for conceptual modeling of signal transmission networks

✓ Design a two IOD signal transmission network suitable for the IOD band

✓ Illustrate the robustness and efficiency of the IOD band design in CeCe simulations

Module 5 allows other modules to use microfluidics devices for their experiments.

✓ Set of microfluidic devices fabricated by PDMS soft-lithography.

✓ Characterization of signal transmission range between wildtype MATa and MATx Saccharomyces cerevisiae cells.

✓ Dynamic characterisation of signal transmission between synthetic MATa and MATx Saccharomyces cerevisiae cells.

✓ Comparison with mathematical model of signal transmission mechanism and estimation of the activation threshold for different cell concentrations.

✓ Demonstrate yeast induced blood agglutination on-chip by human antigen A displayed on cell surface by Yeast Surface Display.