Team:NUS Singapore/Medal Check List

Measurement Modelling Medal Check List
Team Member Attributions
Collaboration
Silver Up Gold Integrated
Overview Methodology Kill Switch for Probiotics Kill Switch for BeeT
Description Interlab Study Parts Experiment Documentation Improvement Demonstrate Safety Future Works
Home

Gold Criteria

  • Integrated Human Practices

    • Based on the interview results, we have a better understanding of the potential challenges in building such kill switch and the importance of having kill switches for different synbio applications. From this understanding, we integrate the knowledge gained by proposing building a framework to make engineering customized kill switch easier.

    Quick Link -> Human Practices
  • Improve a previous part or project

    • Improved the BBa_K116404 phosphate sensor-GFP reporter constructed by NYMU Taipei in 2008. By replacing the weaker RBS BBA_B0032 with a stronger RBS BBa_B0034, the new promoter is more sensitive to phosphate concentration with a higher expression level.

    Quick Link -> Improvement
  • Modelling

    • We have carried out extensive modelling to design the kill switch for probiotics in silico. We built mathematical models of two kill switch designs (one generated by CELLO and one designed by the team) using ODEs. The performance of kill switch designs was evaluated based on the following criteria: number of parts required in a design, metabolic burden on cell, and response/lag time using ADVANCESYN Studio (a design and modeling software). We ran a number of simulations, what-if studies and sensitivity analysis, and identified the sensitive components in the circuit. Results showed that the kill switch design by the team has faster response and lower metabolic burden. Hence, we proceeded to perform experiments with this design. Apart from the kill switch design for probiotics, we also modelled and studied the blue light–pH NAND gate kill switch for BeeT bacteria from team Wageningen_UR iGEM 2016 using the methodology.

    Quick Link -> Model
  • Demonstration

    • We have successfully completed the following tasks:

    • Constructed the temperature-sensitive system with GFP reporter (BBa_K2447014）.
    • Use modelling to facilitate the design of kill switch.
    • Constructed the improved version of the extracellular phosphate sensor with GFP reporter (BBa_K2447000).
    • Constructed the blue light activated inducible system with RFP reporter (BBa_K2447501)
    • Successfully demonstrated that our proposed standardized methodology for designing kill switches was feasible
    • Successfully constructed the plasmid that expressed IM2 protein (anti-toxin) under the control of our phosphate-temperature sensor (BBa_K2447016)
      • Quick Link -> Demonstrate

Silver Criteria

  • Validated Part

    • BBa_K2447013,BBa_K2447014 (IPTG-inducible temperature-sensitive system with GFP reporter)

    BBa_K2447500,BBa_K2447501 (Blue light activated inducible system with RFP reporter)

    Quick Link -> Parts
  • Collaboration
    • Help Stockholm and US AFRL CarrollHS with modelling
    • Meet up and discuss with iGEM NTU, iGEM Peshawar, iGEM Newcastle our projects.
    • Meet up with iGEM York, iGEM Cologne-Dusseldorf, iGEM-NTU, iGEM SVCE Chennai at SynBio 7.0 in Singapore.
      • Quick Link -> Collaborations
  • Human practice
    • Conducted survey and interview with researchers and the public to understand the potential risks related to GMOs, the feasibility of kill switch system and share with them our projects.

    • Conducted interview with the conservationists to understand the potential impacts GMOs on biodiversity.
    • Outreached to member of public about the usefulness of modelling in synthetic biology.
    Quick Link -> Human practice

Bronze