Team:Czech Republic/Goals

Goals


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.

  • Constructed a set of reporter promoters for yeast cells
  • Characterized reporter promoters in all mating types
  • Designed and materialized synthetic MATa and MATx strains
  • Built a synthetic diploid strain with a functional yeast pheromone pathway
  • Demonstrated 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.

  • Constructed a set of yeast plasmids with different mating pheromones and their receptors and contributed to the Registry with 6 BioBricks
  • Verified the correct coupling of the receptors to the yeast pheromone mating pathway
  • Verified the correct expression and secretion of the different pheromones
  • Showed 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.

  • Expressed streptavidin, EpCAM, Anti-EpCAM scFv, c-Myc scFv and anti-HuA scFv on the surface of yeasts
  • Demonstrated the ability of our receptors to bind chosen markers
  • Monitored the dynamic binding of our receptors and their corresponding markers

Module 4 provided modeling and simulation support for other modules.

  • Developed a simulation environment CeCe to capture the complexity of cell-cell signal transmission
  • Designed an IOD chemical reaction network model
  • Developed a schematic architecture for conceptual modeling of signal transmission networks
  • Designed a two IOD signal transmission network suitable for the IOD band
  • Illustrated the robustness and efficiency of the IOD band design in CeCe simulations

Module 5 allowed 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.
  • Demonstrated yeast induced blood agglutination on-chip by human antigen A displayed on cell surface by Yeast Surface Display.