Finding an innovative new topic among all existing projects since the first edition of iGEM is a difficult task which forces us to think out of the box. However, we found out synthetic biology is an endless source of possibilities. Here, we invested in the allergy theme. While there have been previous iGEM efforts about desensitization, allergy detection appeared as a virgin field. We therefore developed a new exciting possibility to detect allergenic predisposition using synthetic biology, via the aggregation of Immunoglobulins E (see Figure 1). For more information on our method, see our Design section.

Figure 1: Process of aggregation. E. coli displaying allergens (orange) and E. coli displaying a DARPin with high affinity for the constant fragment of IgE (blue) agglutinate around the IgE of the patient.

We are certain our wiki content will be useful for the following generation of iGEM teams. We designed it to show everything we learnt during this incredible iGEM adventure and to allow any visitor to reuse knowledge and tools developed during our project. This way, we feel that other teams will be able to create new exciting projects about allergen detection.

Creating a new detection approach leads to designing new protocols. Consequently, a special effort was made to create and test new methods and adapt existing protocols to reach our purpose or simply to control if the behavior of the allergen or DARPin was as planned. This methodologic work required detailed protocols and a special care has been taken for the Protocols page. We feel it would be of great use for the future iGEMers to build on our work.

To reach our purpose, we needed to create a lot of tools, including new parts. We provided all parts to allow the next iGEM teams to build or improve constructions for their own purposes. We designed and validated parts for a robust and constitutive expression of fluorescence in E. coli:

- BBa_K4197014

- BBa K4197013

- BBa K4197012

There are many more than we feel of the greatest interest to produce recombinant allergens and to manipulate antibodies. Visit our Parts page!

We contributed to the improvement of three existing parts on iGEM. We found a new constitutive promoter for E. coli to express fluorescence. We tested them for the red fluorescent protein mScarlet-I (BBa_K2333414) and the blue fluorescent protein mTagBFP (BBa_K592100) with exciting results, in particular for the red fluorescence (Figure 2). The construction with the ihfB800 promoter was used to sort E. coli cells by FACS. We also brought for the OmpA part a crystallographic structure found during our bibliographic research (BBa_K103006).

Figure 2: Second plating of pET-21 b (+)_Ara h 2_OmpA_mScarlet transformed cells (Part BBa_K4197004) and pET-21 b (+)_Ana o 3_OmpA_mScarlet transformed Stellar cells (Part BBa_K4197003). The colonies were selected on LB-ampicillin plates. Pink coloring indicates the expression of the mScarlet.

During this great collaborative project, we contributed to the success of the Interlab by giving them feedback to clarify the proposed protocol. Then we calibrated our plate reader and conducted the Experiment 1, and gave our results to iGEM (click here for the Excel calibration and the Experiment 1 results sheets). Since our plate reader was new and dedicated to students from our Institute, it has been also a rewarding experience for us since it allowed us to take charge and set up this new device.

Our modeling effort was thought to serve directly the design of experiments by studying molecular interactions between IgE, DARPin and Allergens. Then, a second model linking state-of-the-art fluid mechanics and biology was conceived as a pioneering model for the iGEM community to study interactions between bacteria and predict aggregation (see Figure 3). Our effort illustrates how modeling strategy could be central to improve the wet lab set-up. More details are in the Model section.

Figure 3. General overview of the modeling strategy. The model is constituted of two parts : a complexation model and an aggregation model. Each model allows us to approve the feasibility of the project, optimizing the concentration, making aggregation assays and is useful for our cytometry strategy.

Ethics are a crucial aspect of sciences, and even more in health-related projects. This is why we engage in an ethical approach from the start of this project (see Figure 4). These led to deep thoughts nourished by exchanges with both stakeholders of allergy detection and specialists for each step of the project. This reasoning was shown and summed up in our Integrated Human Practices page. Our approach provides a demonstration of the application of ethics on an engineering project.

Figure 4: illustration of the ethical approach

We built our whole Education strategy around the central value of accessibility: to anyone, anytime, and anywhere. From pedagogic studies and classification of communication supports, we opted for two main actions. The first is the creation and use of bacteria and immune cell models to explain synthetic biology and allergies whatever the scientific background (see Figure 5). The second is a whole collection of videos named "Cracking Allergies" available to all online. They address every question we receive about allergies from the general public during our participation in scientific exhibitions and meetings. Both supports were used in subsequent educational efforts such as media communication and interventions in high school. The success we had throughout these events as well as online has proved to us that a well-constructed educational effort is very rewarding and we provide all the details on our wiki so that future iGEMers can build upon our material and experiences. Go to our YouTube channel or iGEM Universe for videos and the Education and Communication wiki to know more about it !

Figure 5: Bacteria models to explain the DAISY project

Meetups are the best way for the iGEM community to grow and for the research community to share ideas, to find new ways for self-improvement and to have feedback about your project. However, organizing a meet-up represents a great investment, and requires a whole array of competences. Teams from Münster, Hamburg, and Toulouse have organized meetups in person this year (see Figure 6).

Figure 6: Group picture of the students

We gathered to think about all the issues we faced and decided to co-create and share a Meet-up guide for all the next iGEM teams. In this document, we share and comment our experiences and provide many tips to help organize an in-person meetup and avoid mistakes. The document is available on our three wikis and we also want it to be uploaded on the iGEM community meetup page. Of course, we hope that thanks to our guide, the upcoming meetups organization will be facilitated.. More details are available on our Collaboration page.