Team:Hong Kong HKU/Engineering

HKU EVOLVE - Teams

ENGINEERING ADVANCES



Through our iGEM project, we have demonstrated the proof-of-concept of our novel Cre-loxp circuit through computational modelling and showed that our system can produce proteins in a defined ratio with high stability. We adopted the engineering cycle to perfect our system through the process as follows:

Research: Biomanufacturing industry has a need to produce proteins at defined ratios. Research on microbial bioprocessing advances. (See more...)

Imagine: Can we use a single microorganism that can express two or more phenotypes for different protein production?

Design: Develop the cre-loxp circuit design. (See more...)

Build: Construct the computational model to simulate the cell growth kinetics and phenotype switching dynamics. (See more...)

Test: Run the model and obtain the results. (See more...)

Learn: Realised that our system can produce phenotypes at a constant ratio in the long run with very low variations. (See more...)

Improve: Release the assumptions in the computational model (See more...)

As we do not have full access to laboratory due to the COVID situation, we mainly put our focus on modelling and program development.

With limited time for wet lab, we were able to:

  • Design parts

  • Amplification of parts

  • 5 Site-directed mutagenesis on Cre recombinase

  • Rounds of Lox-reporter construct and Cre construct assembly

  • Co-transformation and pilot experiment

  • In vitro Wild-Type Cre shuffling

  • Multiple rounds of In vivo Wild-Type Cre shuffling with optimisation of parameters

    • Learn more about our models and program by clicking on the cards below!