Difference between revisions of "Team:Newcastle/MedalRequirements"

• BBa_K2205002 - J23100-deGFP
  • We have BioBrick standardised the deGFP Green Fluorescent Protein (GFP) variant for the first time. We created an expression construct (J23100 promoter and B0034 RBS in front of deGFP) and characterised the deGFP. Check the registry page or our results page to see full experimental data.
  • This part was made to be one of the potential reporter modules in our multicellular biosensor framework.
• BBa_K2205004 - T7-Sarcosine Oxidase
  • We have BioBrick standardised the sarcosine oxidase enzyme for the first time. We created an expression construct (T7 promoter and B0034 RBS in front of SOx CDS) and demonstrated that SOx is capable of catalysing the reaction of sarcosine to formaldehyde. Check the registry page or our results page to see full experimental data.
  • This part was made to be one of the potential adapter modules in our multicellular biosensor framework.
• BBa_K2205009, BBa_K2205012, and BBa_K2205015 - Sensynova Multicellular biosensor Framework Modules
  • We have designed, standardised, and characterised three modules (IPTG detector, blank processor, and sfGFP reporter) to act as a proof-of-concept for our multicellular biosensor framework. We demonstrated that each module is able to accept its input molecule, and produce its output molecule as a result. Check the registry pages (BBa_K2205009, BBa_K2205012, and BBa_K2205015), or our results page to see full experimental data.

At the beginning of iGEM, we created a flyer showcasing the strengths of our team and the various ways we could help other teams. As a result of this, we were contacted by three teams: Edinburgh overgraduate team, Exeter, and Evry Paris-Saclay. For the Edinburgh team, we used our modelling knowledge to help fix their model. We aided the Exeter team by repeating one of their experiments, and they gathered single-cell fluorescence data for our deGFP construct using their FACS machine. Finally, Paris designed a novel psicose-regulated promoter to be used in a biosensor. We then incorporated their promoter into our framework so that many different variants of a psicose biosensor could be designed, made and tested.

Check out our collaboration page for more information

Our human practises work has focused on addressing technology uptake, which is one of the challenges that we identified to biosensor development and deployment. This took place in three main stages. First, we determined the current state of dialogue by consulting previous dialogue studies and reviewing how language is used in the media. We then generated our own guidelines to help future researchers to develop dialogue in a constructive way. Finally, we put this into practise by creating activities and sharing our work in a way that established a dialogue and encouraged discussion.
Check out our Human Practices (Silver) page for more information.

Sensynova was a project founded in Human Practices. Each branch of our project is rooted in a barrier to biosensor implementation identified by in-depth conversations with stakeholders. We identified not only technical issues, but also societal issues, as these are equally important in ensuring successful implementation of biosensor projects. The aims of our human practices were to:

1. - Determine how current biosensor developers design, produce and implement biosensor projects
2. - Use these talks to determine the common barriers to biosensor development and implementation
3. - Create novel solutions to identified problems, producing frameworks and guidelines to aid future researchers

To achieve these aims, we emailed, skyped and attended conferences to speak to stakeholders in biosensor development, from the early research stage to the end-user. Check out our integrated human practices page for more information

BBa_K2205026 - pSB1A51 (Promoter Library Screening Plasmid) - Improvement over BBa_J61002

• Transcriptional insulation added before promoter cloning site (PCS)
• lac operator added after PCS for additional characterisation ability
• RBS cloning site added to allow testing of promoters with any desired RBS
• mRFP1 CDS replaced with the CDS for the more fluorescent sfGFP
• Promoters can easily be removed from pSB1A51 and assembled into another plasmid/with another BioBrick using BioBrick assembly

BBa_K2205005 - Fim Standby Switch - Improvement over BBa_K1632007

• Replaced E0040 CDS with eforRed chromoprotein CDS to allow easy visualisation with the naked eye
• Added RhlI CDS on the reverse strand upstream of the flipable promoter to allow modular addition of secondary reporter CDSs (can be added within same cell, or in a different cell of a multicellular communiuty)
• Provides a 'dual reporter' which produces eforRed in its 'default' state (i.e. no signal detected), and a protein of choice in its 'on' state (i.e. when a signal is detected)

For our project, we built three types of models. The first was an agent-based model which simulated our multicellular biosensor framework. This model gave insight into the optimal ratio of cell-types to have in the system. This information was used during experimental characterisation to optimise our system. Our second model was a statistical, multifactorial Design of Experiments (DoE) approach towards optimising Cell-Free Protein Synthesis (CFPS) systems. This statistical model was used to generate an experimental design to gather data on the importance of certain supplements in CFPS systems, and then use the experimental data to optimise CFPS systems.
Find out more about our models here.

We have produced a functional multicellular biosensor based on bacterial quorum sensing. Each part has been tested and characterised singularly. The co-culture of the 3 bacterial types are proved to form a working biosensing device. More details can be found here.