Team:Sheffield/Medals

A template page

MEDALS

1. Register and Attend

We have registered and plan to attend the Jamboree

2. Deliverables

  1. Team Wiki

    Our wiki can be found at 2016.igem.org/Team:Sheffield.

  2. Poster

    We will be attending the Jamboree, and defending our poster

  3. Presentation

    We will be attending the Jamboree, and presenting our project.

  4. Project Attributions

    The attributions can be found at 2016.igem.org/Team:Sheffield/Attributions.

  5. Registry Part Pages

    All of our submitted BioBricks can be found at 2016.igem.org/Team:Sheffield/Parts

  6. Sample Submission

    We will be sending off our BioBricks by the official deadline.

  7. Safety Forms

    We have already submitted all the necessary safety forms by the appropriate deadline.

  8. Judging Form

    We have completed and submitted the judging form by the deadline.

3.Attribution

The attributions can be found at 2016.igem.org/Team:Sheffield/Attributions.

4.Document at least one new standard BioBrick Part or Device used in your project/central to your project, and submit this part to the iGEM Registry.

For our project we came up with two reporter ideas for our diagnostics device. The first plan was to use Hemerythrin. Hemerythrin when bound to iron, in the presence of oxygen, undergoes a conformational change. That change results in a colour change from colourless to pink. Our team tried Hemerythrins from three different species. Mc Hemerythrin (BBa_K2016003) and Td Hemerythrin (BBa_K2016004) will be submitted to the registry.

1.Experimentally validate that at least one new BioBrick Part or Device of your own design and construction works as expected. Document the characterization of this part in the Main Page section of the Registry entry for that Part/Device. This working part must be different from the part you documented in Bronze medal criterion #6.

The second reporter idea was attaching a ryhB binding sequence to GFP. In the absence of iron, ryhB siRNA binds to ryhB binding site and marks GFP mRNA for degradation. In the presence of iron, the Fur regulator represses ryhB, and GFP is fully translated. We used two different promoters from the Registry (BBa_J23100, BBa_J23106) to drive expression of our ryhB-GFP constructs. Our team then used fluorimetry to investigate GFP expression in the presence and absence of iron. This data experimentally validated that GFP expression works as expected under the different strength promoters. These results can be found on the ryhB-GFP Registry page (BBa_K2016005), as well as on our wiki https://2016.igem.org/Team:Sheffield/Parts. All of our BioBricks are in the pSB1C3 plasmid, and follow all the safety standards. This data also validates the expression strength of the two Berkeley 2006 promoters.

2.Collaboration

A diary of all of our collaborations can be found at 2016.igem.org/Team:Sheffield/Collaborations.

We supported iGEM teams Evry and Ionis-Paris, by attending their European iGEM mini Jamboree in Paris. This big pool of European teams that we all helped create was an amazing opportunity to exchange project ideas, and prepare for Boston. We also joined the Westminster team in their UK meet up in London. In addition, we helped Warwick at their open day at University by visiting them, and providing an activities stall.

Having an iGEM alumni advisor whose project was on Cystic Fibrosis, we joined team Tel Hai (Israel) in their 65 Roses campaign to raise CF awareness. We also contributed to the “Our iGEM experience” video by TecCEM, along with 4 other countries. Finally, we completed three surveys for teams Aachen, Paris-Saclay, and Groningen.

3.Demonstrate how your team has identified, investigated and addressed one or more of the Human Practises issues in the context of your project

On the Sheffield 2016 wiki you can find an interactive timeline that narrates the story of our Policy & Practises investigation, and how the different milestones have had an impact on the design and execution of our project.

2016.igem.org/Team:Sheffield/Integrated_Practices

While brainstorming, as most of the expertise of the team leaned towards Microbiology, this led the team to decide the antibiotic resistance project. The first chapter of our story begins with the quest to answer the question, with the help of Dr Wainwright, of how we ended up in a near post-antibiotic era. The next step was to gain a better understanding of how many stakeholders were involved in this complex matter of resistance with the help of a politics lecturer. Moreover, during the early steps of designing a device prototype, the different end users were explored by approaching hospitals, doctors, patients, and government bodies.

The team also organised two mini iGEM competitions for 17-year-old students in high schools. The students were invited to a fair where we presented posters of previous iGEM projects. They were then divided up in teams with 1-2 iGEMmers to guide their brainstorming. In the end, advisors acted as judges, and gave out the prizes.

1.Expand on your silver medal Human Practices activity by demonstrating how you have integrated the investigated issues into the design and/or execution of your project.

For our Human Practises the team approached several people to obtain feedback, and return to the project drawing board for improvements. To begin with, to better understand the problem, Dr Simon Rushton, a politics lecturer, helped us identify potential stakeholders. Dave Ogelsby is in the UK government Rapid Diagnostics Steering Committee that surveys implementation of the antimicrobial resistance five-year strategy plan. He pointed out that our device would help prevent doctors prescribing antibiotics to viral infections. In order to understand the doctor and hospital end users, we talked to Mrs Thompson. We informed modelling that our test needed to be shorter than 2hours, and our device needed to use a prick needle for sampling. This is because blood tests are only allowed in lab hospitals. For the full story of our Practise, and how they have influenced the design of our project, please visit: 2016.igem.org/Team:Sheffield/Integrated_Practices

2.Improve the function OR characterization of a previously existing BioBrick Part or Device (created by another team, or by your own team in in a previous year of iGEM), and enter this information in the part's page on the Registry.

Sheffield 2016 has improved the function of the BBa_J23100 part, by adding an RBS site downstream of the promoter (BBa_K2016005). BBa_J23100 was originally submitted to the Registry by Berkeley 2006. Our team has also improved the characterisation of both BBa_J23100 and BBa_J23106. These parts are a strong and medium promoter respectively, that we have used to design our iron detecting device. We have experimentally validated through fluorimetry that there is indeed a significant difference between expression levels of GFP coupled to the strong and medium promoters. Comparative analysis of promoter strengths can be directly interpreted from the data we obtained. This data can be found both on the original part experience pages of BBa_J23100 and BBa_J23106, as well as at 2016.igem.org/Team:Sheffield/Parts.

3.Demonstrate a functional proof of concept of your project. Your proof of concept must consist of a BioBrick device.

We have designed a device (BBa_K2016005) using the BBa_J23100 promoter, adding an RBS site to it, and coupling it to a GFP with a ryhB binding sequence.

In a bacterial infection, lipocalin is produced by the human or animal organism. Lipocalin prevents bacteria from uptaking iron. We use GFP expression as a means to determine and quantify the presence of bacterial infection, by utilising the ryhB pathway.

In the presence of lipocalin (thus, absence of intracellular iron), ryhB can freely degrade GFP mRNA in bacteria. In the absence of lipocalin (thus, presence of intracellular iron), the Fur regulator represses ryhB, and GFP is produced.

We have used fluorimetry to experimentally validate that our engineered bacteria change their levels of GFP expression after addition of a defined amount of iron.