Bronze Medal

As we met all the deadlines and created separate pages for Competition Deliverables,Project Attributions, Project Description and Contribution, we think we are eligible for the Bronze medal.

Competition Deliverables
We believe we meet this criteria because we have completed the following:
- Wiki (you are browsing this now!)
- Project Promotion Video
Project Attributions:
Check out our team’s attribution page
Project Description:
Over the next 35 years, the world’s growing population will demand more food than has ever been produced in human history. Fungal pathogens constitute 75% of all plant infections, and current solutions are falling short, either unable to keep up with rapidly mutating pathogens or failing to deploy in the most effective time window. In response, we have developed Sporadicate, a broad-spectrum biofungicide consisting of a B. subtilis spore system that eliminates the time lag between diagnosis and treatment. By engineering proteins on the outer coat of the spore, we enable the timely detection of fungal pathogens, with selective germination into vegetative cells exhibiting biocontrol properties, as a response. Given the innate durable properties of spores, our system would be robust, easily storable, widely applicable, and cost-effective. Meeting the needs of farmers across the world, especially in resource-poor areas where vulnerability to phytopathogens is greatest, with far-reaching consequences. Check out our team’s project description
Contribution:
We are the first team to assemble parts BBa_K4212034 and BBa_K4212038. We are also the first team to generate B.subtilis germinant receptor chimeras: part BBa_K4212050 and part BBa_K4212051 by cloning in E.coli. To support groups attempting to alter binding specificity of a given protein, we have developed a computationally efficient tool to sample and test in-silico binding affinities between a ligand and mutants within the vast combinatorial space given a set number of residues to be mutated. We carried out extensive Human Practices work, including development of a product risk assessment framework, which we believe will aid future teams dealing with plant fungal pathogens or working with B.subtilis.Check out our team’s contributions

Silver Medal

We have also met all the criteria for silver medal. We have demonstrated engineering success,collaboration, human practices, and proposed implementation.

Engineering Success:
During the initial design phase we relied on a variety of computational tools (such as Benchling, GeneArt, Alphafold and Molecular dynamics simulations) to inform the construction of our chitinase display strategy and our take on a self-digesting circuit for biocontainment. Furthermore, not only did we build (and troubleshoot) these components, we also discovered strengths and weaknesses of the Subtitoolkit, the MoClo collection of parts we have used across our project to engineer bacillus. With Sporadicate, both computational and laboratory work were linked in continuous yet intimate engineering cycles. In order to showcase how the engineering design-build-test-learn cycle was successfully adopted across our project, we have chosen two examples: the development of a fusion protein system for surface display on spores in the wet-lab, and the establishment and execution of a strategy for germinant receptor engineering in the dry-lab. Please see our engineering success page for more details. Check out our engineering success
Collaboration:
Aiming to provide an interactive platform for iGEMers, researchers, and the general public interested in synthetic biology, we organised the 2022 UK-wide iGEM Meetup and hosted around 50 participants in-person and online. The event spanned two days, the first consisting of seminars and Q&A sessions with leading researchers in the synbio space working on applications from artificial cells to biosynthesis in resource-poor regions. On the second, a mock jamboree with five UK teams, and an expert panel of judges including iGEM alumni and an ambassador who provided personalized feedback to each was hosted. We also participated in the 2022 Agricultural iGEM teams meetup. We presented our idea and the work we had done so far for the different components of our project. We engaged with other teams, discussing challenges and successes experienced and shared advice about project ideas. Check out our collaborations page here.
Human Practices:
Our idea development and implementation plan have been consistently informed by stakeholder interviews with our final users, farming union representatives, agricultural experts as well as experts in synthetic biology and policy. Through the innate durability of spores, our product has been designed with accessibility in mind. This means we can effectively address the threat of fungal diseases in the regions of the world most at risk, with a tool that can be easily adopted by subsistence and smallholder farmers who carry the majority of the burden of wheat production there. Percentage yield losses are higher in such areas due to the difficulties in the implementation of existing solutions such as expensive novel resistant cultivars. Our project can mitigate the impact of pathogenic fungi by being non-race specific and applicable to different crop varieties, with no reduction in efficacy. Learn more about our approach on our page! Check out our human practices page here.
Proposed Implementation:
will initially target smallholder farmers who are more urgently in need of a cheaper solution. Smallholder farmers will only require a smaller quantity of our product making them a useful customer whilst the scale of our manufacture is increasing. We will then seek to raise money for large scale manufacturing facilities based on our success in our beachhead market. We will first seek regulatory approval in the US, as many countries follow their lead for food safety and environmental regulations. Although we have designed our solution to be compliant with European regulations, we anticipate it being more difficult to obtain approval as well as public acceptance of our product in the EU than in other parts of the world, given the results from our public outreach efforts. Therefore, we will not aim to sell our product in the EU until we have secured a substantial market share elsewhere.Check out our implementation page here.

Gold Medal

Finally, combining all the effort from different aspects of our project, we think that we are eligible for the gold medal. We have met 4 out of 6 eligibilities of the gold medal criteria.
‍

Integrated Human Practices:
- Combining all the views we gatheredfrom farmers, stakeholders, and the public, we sensed the necessity to buildboth temporary and root cause approaches like ours, since there is nogovernmental intervention or regulations in solving the crisis of foodinsecurity.
- We also learned that the timeframeto modify the wheat genome to be fungal resistant is especially long. That iswhy we decided to use spores as a fungicide platform.
- Both our wet lab experiments and drylab modeling got feedback from our integrated Human Practices. Check out more on our integrated human practices page.
Project Modeling:
To inform wet-lab experiments in thedirected evolution aspect of the project, our dry lab team investigated thestructure and behaviour of the GerAB receptor, informed the design of ourfusion protein linkers, and inferred interaction parameters between differenttaxa present in the wheat rhizosphere. Check out more on our modeling page
Proof of Concept:
- We would like to develop our biofungicide into a spray, both for easy and timely application.
- Based on our research, we found outthat the wild-type B.subtilis exhibits anti-fungal activities.
- We proved that wild-type B.subtiliscan inhibit fungal growth. By doing that, we can also see the different extentof fungal inhibition activities B.subtilis has based on their dilutionratios. Check out our proof of concept page for more information.
Partnership:
- We collaborated with Paris/Bettencourt and Costa Rica iGEM teams. Check out more on our partnership page

Our Goal: Special Prizes

Best Integrated Human Practices:
Food is an integral part of human existence, and as such is a sensitive issue. Food insecurity is a challenge that affects every community around the world, most acutely those part of marginalised groups. The agricultural industry at a global scale is also characterised by inequality, whether that be environmental, economic, technological or knowledge based, with resource-poor farmers competing against highly industrialised enterprises. Given this landscape, we wanted to ensure we were engaging in responsible innovation, acknowledging that innovation can have unpredictable outcomes and raise questions and dilemmas. During our ideation phase we went through several cycles of the Double Diamond framework as we optimised our idea based on feedback garnered from a diverse array of stakeholders spanning key groups: farmers (our final users), experts, regulatory bodies and the general public. To better understand the impact our research could have, beneficial or otherwise, we adopted the UKRI AREA framework.
‍
Best Model:
To inform wet-lab experiments in the directed evolution aspect of theproject, we investigated the structure and behaviour of the GerAB receptor. Weinitially modelled the structure of the complex. Subunit interaction sitesidentified were used in designing the chimeric receptors tested in the wet-lab.Key residues involved in binding were identified using a range of tools, andinformed the creation of a mutant library to be tested in future research.Modelling also informed the design of our fusion protein linkers. A range ofdesigns with parameters varied according to research were explored, informingthe selection of anchor protein as well as length and composition of thelinker. To assess the effect of our bio-fungicide on the wheat soil microbiome, we modeled the soil microbiome as a generalised Lotka-Volterra system with random community interactionsrends. This was used to make predictions regarding the behaviour of our spore system.
‍
Best Software Tool:
To support groups attempting to alter binding specificity of a given protein we have developed a computationally efficient tool, InFinity 1.0, to sample and test in silico binding affinities between a ligand and mutants within the vast combinatorial space given a set number of residues to be mutated. Our tool is an open-source framework that can be applied in re-egineering any protein to alter and improve ligand specificity and affinity. It not only offers researchers viable mutants but also provides ranking and multiple sequence alignments of mutants to provide a better understanding of what types of mutations are more likely to cause positive change in ligand affinity. Although in early development, we have found our tool comparable to other established closed source tools, and a viable aid as the first step in in silico rational design.
‍
Best Supporting Entrepreneurship:
Firstly, interested in knowing the potential of our Sporadicate project in the future fungicide market better, we made a thorough market analysis. It assists us in perceiving the growing market for fungicide, as the changing climate promotes the spread of fungal infections, and growing population poses a pressure to increasing food production. Thus, we proceeded with a manufacturing and cost analysis, which lays out the manufacturing technology, material cost, packaging and delivering cost, and market regulations. This concluded the economic feasibility of manufacturing our project and pushing it to the market. Considering the current major players in fungicide market, we did a competitor analysis, comparing different aspects of our project with other major fungicide products. Our product shows a timely application with lower cost and labour treatment, non-toxicity towards environment, and improvement of well-being for both farmers and consumers.
‍
Best Sustainable Development Impact:
Due to the rapidly changing environment, the frequent mutations of fungi, and the time-consuming scheme of cultivating resistant crops, traditional diagnostic and treatment techniques fail to prevent the rampant spread of fungal diseases, a challenge to resolve if we are to achieve Goal (#2) Zero Hunger .Our spore system can quickly detect and eliminate the fungal pathogens, minimizing the loss of crop yields. With fewer applications of our spores, we help farmers to reduce expense on purchasing fungicides, while avoid contaminating the environment. By developing a platform with B.subtilis spores displaying chitinase on the surface, we constructed a non-toxic plant disease control system contributing to Goal (#3) Good Health and Well-being, and Goal (#9) Industry, innovation and infrastructure of SDGs. Our self-digesting plasmid also ensures the spore is free of foreign DNA, thus meeting the expectation of Goal (#12) Responsible Consumption and Production.