Along our iGEM journey, the team has set out to design a living therapeutic platform based on engineered probiotics for asthma treatment. Asthma is one of the most prevalent chronic respiratory diseases across the globe, with millions of patients taking daily medication that often does not manage symptoms, usually not addressing the root cause of the inflammation. Our project proposes a new strategy for biological delivery: using Lactobacillus plantarum, a safe probiotic harboring natural anti-inflammatory activity, engineered to deliver therapeutic RNA molecules directly to inflamed lung tissue [1, 2, 3].
Even from the beginning, our focus was not only on asthma but also on creating a modular platform that can be utilized for many pulmonary and gastrointestinal diseases. By sharing the design decisions along with any obstacles and solutions with the iGEM community, we hope to shed light on the path for future iGEM teams interested in synthetic probiotics and RNA therapeutics [4].
To bring this vision to life, we created multiple new basic and composite parts that can be used for various applications. All of them are listed on our Parts Page. Some of our most impactful parts are:
In our project, this part allows efficient Loading of therapeutic CO-BERA into bacterial membrane vesicles and delivers it to Lung epithelial cells. We believe that this part represents not only a solution for our project but also a new building block for the iGEM community, a step toward programmable probiotics with precise molecular delivery capabilities [5, 6, 7].
The potential of this composite part goes far beyond our application:
Modular RNA loading system: DUF4811–L7Ae BBa_25BFTYLX can be adapted to load different RNA scaffolds, making it useful for diverse RNA-based therapies [6].
Synthetic biology toolbox expansion: By linking RNA-binding proteins to transmembrane anchors, this design strategy provides a generalizable way to spatially localize RNA inside bacteria [8].
Therapeutic delivery: Future iGEM teams can reuse or modify this composite part to deliver RNAs, proteins, or other molecules via bacterial vesicles [9, 10].
CO-BERAs BBa_250VF2PY are highly stable RNA scaffolds that can simultaneously deliver multiple siRNAs. In our project, CO-BERA was tailored to silence TSLP for asthma treatment, but the system itself is broadly applicable. Our team expanded the application of the CO-BERA system by integrating it into a probiotic delivery platform [11, 12].
CO-BERA (BBa_250VF2PY) use goes beyond asthma as it orean be repurposed to target cancer drivers, viral infections, autoimmune pathways, or metabolic diseases. Future iGEM teams can adopt our composite design to load their own siRNA sequences into CO-BERA, achieving precision. We provide the community with a versatile RNA-based therapeutic chassis that can serve as a general platform for synthetic biology applications [13, 14].
In addition to designing new parts, we also worked on improving existing biological tools. One of the challenges in therapeutic delivery is ensuring that cargo such as RNA can escape the endosome after entering host cells. Natural Listeriolysin O (LLO) has this property, but its high virulence makes it unsuitable for therapeutic applications [15].
To overcome this, we engineered a mutated LLO variant BBa_25P1C1BV with reduced pathogenicity compared to the wild-type protein, while preserving its ability to mediate endosomal escape [16].
Why this matters: Endosomal escape is a major bottleneck in RNA therapy; only ~0.3% of internalized RNA typically escapes into the cytoplasm [17].
Our improvement: By modifying LLO, we provide a safer and more controllable tool for enabling RNA delivery without compromising host safety [16].
Impact: This improvement makes LLO compatible with probiotic-based therapeutics and provides a reusable part for the iGEM community interested in safe intracellular delivery systems [18].
We believe that this improved LLO part will be valuable beyond our project, enabling applications in RNA therapeutics, vaccine delivery, and engineered microbial therapies [18].
Safety is one of the fundamental considerations in designing our project. While working on the innovative therapeutic platform employing Lactobacillus Plantarum, biosafety was equally important as proving the functionality. Engineered microbes can do wonders for medicine and biotechnology, but they can be dangerous if released into any unintended environment or if they transfer their genes to other organisms [19].
To qualify the chassis to meet these requirements, our team incorporated multi-layer safety mechanisms, namely toxin-antitoxin kill switches and environmental sensors. These safety features serve to:
1. Prevent survival of engineered bacteria outside the intended site (lungs) [20, 21, 22].
PhoB Promoter (phosphate sensor): Ensures that bacteria cannot survive in the bloodstream, where phosphate levels are high [20].
Thermosensor RNA: Detects temperature drop outside the host and kills the bacteria to avoid survival in the external environment [22, 23].
2. Reduce the risk of horizontal gene transfer (HGT). One of the major biosafety concerns with GMOs (genetically modified organisms) is gene transfer to other microbes. By ensuring rapid elimination of engineered L. plantarum under non-permissive conditions, our system significantly reduces the window of opportunity for HGT [24] for more details see as mentioned in the Safety page.
Our multi-layered system generally offers a strategy that can be readapted by other iGEM teams. These tools take safety to a higher level without interference with the applications, whether for therapeutic probiotics, environmental biosensors, or biocontainment of industrial strains [25].
By editing and sharing these designs, we hope to contribute some practical biosafety tools to the iGEM community so that future teams can consider safety issues in their therapeutic and environmental projects [19].
Many iGEM teams use freeze-drying to prepare engineered probiotics as a stable powder for transport and storage. However, when bacteria are equipped with thermosensor-based kill switches, the sharp temperature decrease during freeze-drying can mistakenly trigger cell death. This creates a conflict as the method needed to formulate the probiotic also eliminates the engineered strain [26].
To address this, we designed a freeze-drying preparation protocol that prevents accidental activation of the thermosensor-linked toxin–antitoxin system during manufacturing. This protocol allows engineered bacteria to survive freeze-drying while preserving the kill switch's full function after rehydration in the host or exposure to non-permissive environments [27].
Problem-Specific Protocol – We provide the first documentation addressing the conflict between freeze-drying and thermosensor-based kill switches [28].
Generalizable Tool – Future iGEM teams developing safety circuits triggered by temperature can adapt our strategy to ensure their bacteria remain viable during preparation but safe in the environment [27].
Practical Resource – We share a protocol PDF that outlines design considerations and a workflow for integrating freeze-drying with thermosensor-equipped microbes.
How This Helps the Community
1. Teams can confidently apply freeze-drying without losing engineered strains prematurely.
2. Safety circuits remain reliable, preventing accidental survival outside intended conditions.
3. Our documentation saves time by providing a ready-to-use framework for others facing the same problem.
As our team successfully designed and optimized a dry powder inhaler (DPI) by integrating the most preferred parameters reported across multiple DPI types. This allowed us to minimize airflow resistance (≈0.022 kPa^0.5·L⁻¹·min⁻¹) while maximizing aerosolization efficiency (MMAD ≈ 3.0 µm, FPF > 60%, ED > 85%). By combining CAD modeling, CFD simulations, and prototype testing, we designed a device that is both high-performing and manufacturable with 3D printing and scalable injection molding.
Even though the primary application for this device was to administer engineered Lactobacillus plantarum, its uses certainly go beyond just probiotics. Its modular design and the optimized parameters set make it fit for delivering diverse therapeutic powders, ranging from biomolecules to peptides to engineered microorganisms.
Our results demonstrate that the platform achieves reproducible particle dispersion with minimal dose loss. In addition, maintaining patient usability through clinically acceptable resistance levels. The integration of numerical simulation with experimental validation reduced development cycles and provided a strong scientific foundation for design improvements.
This hardware contributes not only as a delivery platform for our project, but also as a general-purpose, adaptable DPI framework for future iGEM teams and biomedical applications. Moving forward, cascade impactor studies, microbial viability assays, and usability testing will further advance this engineered prototype toward real-world clinical translation.
Our project introduces PULMORA, an integrated multi-scale mathematical framework that supports the design, validation, and translation of probiotic-based therapeutics. Beyond guiding our own project, PULMORA was built as a reusable modeling toolbox for the iGEM community [29].
We put together visualization Plots such as sensitivity analyses, Monte Carlo simulations, and contour plots to help translate complex modeling outputs for both technical and non-technical users [33].
This assists teams not only in making models but also in communicating their results on the wiki and on disseminating them to the community [33].
Modular Design: Each Pulmora model has the potential to be used on its own or to be integrated into a larger pipeline [29].
Transparency: All assumptions, equations, and code are documented openly for modification [30].
There are many diseases caused by the accumulation of specific proteins. Any team worldwide can use our model equations, parameters, and code to validate knockdown of the causative protein — although they must design siRNAs specific to the targeted mRNA [13].
Furthermore, PULMORA is a community resource: it enables iGEM teams to validate gene circuits, predict therapeutic efficacy, and optimize manufacturing processes, helping convert theoretical designs into real, testable solutions [29].
It allows iGEM teams to validate their gene circuits, predict therapeutic efficiency, and even optimize manufacturing processes, converting theoretical designs into real, testable solutions. With our reusable models the therapeutic world can have a new era of high validated high specific treatments for the incurable diseases [29].
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Stankovic M, Veljovic K, Popovic N, et al. Lactobacillus brevis BGZLS10-17 and L. plantarum BGPKM22 exhibit anti-inflammatory effects by attenuating NF-κB and MAPK signaling in human bronchial epithelial cells. Int J Mol Sci. 2022;23(10):5547. doi:10.3390/ijms23105547.
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Our journey was filled with a lot of activities and interactions in two main directions: asthma and synthetic biology. We aimed to reshape the community awareness and perception toward these topics. Tailoring materials and activities appropriate to the targeted age group was the milestone of planning the trip. In the following we represent materials that we create to achieve our goals.
1. Our team came up with a creative way to teach kids about asthma: we wrote two simple, catchy songs that make the topic easy to remember. The lyrics explain the common symptoms, how the inhaler helps, the right way to use it, and what to do when signs of asthma appear. After writing them, we turned the lyrics into full songs with recorded voices using AI tools, making them more engaging and fun to listen to.
2. We created a simple illustrated story for children that follows a young boy through his day as he experiences an asthma attack. The story shows what triggers he faces and how the situation is handled, all in a way that's easy for kids to understand.
3. We also designed a fun mobile game " as mentioned in Education "that works for both kids and adults. First game main goal is to raise awareness about the common triggers of asthma in an engaging and entertaining way. The other game goal is providing knowledge of medical terms regarding asthma & the respiratory tract in general.
4. Since asthma is more common in children and because they spend most of their time at home under the care of their parents, we developed a set of materials tailored for families. These included an illustrated storybook for kids to help them recognize common asthma triggers in their daily lives, a guidebook for parents explaining the correct ways to manage asthma attacks, and a poster summarizing key tips on handling pet-related triggers at home.
5. We prepared a guidebook that introduces asthma, its symptoms, and the treatments currently used, along with simple daily tips to reduce exposure to triggers. It was designed in an easy-to-read style with clear illustrations to make the information accessible to everyone.
6. We created a public podcast that introduces asthma, its symptoms, and how to manage it in a simple and accessible way. By avoiding medical jargon and using everyday language, we ensured the information could reach and resonate with a wider audience.
7. The team designed and released three different posters throughout the journey, each targeting the general public and raising awareness in a specific area. One poster explained the correct way to use an asthma inhaler, another highlighted the general health risks of smoking, and the third focused on cleaners, showing them how to use personal protective equipment to stay safe from the materials they work with.
1. We produced a short social media video as an introduction to synthetic biology and its various applications in society. The goal was to capture people's attention, spark their curiosity, and raise awareness about this growing field.
2. Our team created a guidebook that introduces the basic principles of synthetic biology, its key applications, and the main tools used in the field. It was designed to give the public a clearer understanding with more scientific detail, while also providing additional educational resources at the end for those who want to keep learning.
Garner K. L. (2021). Principles of synthetic biology. Essays in biochemistry, 65(5), 791–811. https://doi.org/10.1042/EBC20200059
Wang, Y. H., Wei, K. Y., & Smolke, C. D. (2013). Synthetic biology: advancing the design of diverse genetic systems. Annual review of chemical and biomolecular engineering, 4, 69–102. https://doi.org/10.1146/annurev-chembioeng-061312-103351
Yang, L., & Lu, Q. (2025). Beyond Cutting: CRISPR-Driven Synthetic Biology Toolkit for Next-Generation Microalgal Metabolic Engineering. International journal of molecular sciences, 26(15), 7470. https://doi.org/10.3390/ijms26157470
Kumaran, A., Jude Serpes, N., Gupta, T., James, A., Sharma, A., Kumar, D., Nagraik, R., Kumar, V., & Pandey, S. (2023). Advancements in CRISPR-Based Biosensing for Next-Gen Point of Care Diagnostic Application. Biosensors, 13(2), 202. https://doi.org/10.3390/bios13020202
Zhao, M., Tanourlouee, S. B., McCracken, S., & Williams, P. R. (2025). Genetically encoded biosensors of metabolic function for the study of neurodegeneration, a review and perspective. Neurophotonics, 12(Suppl 2), S22805. https://doi.org/10.1117/1.NPh.12.S2.S22805
Burgos-Morales, O., Gueye, M., Lacombe, L., Nowak, C., Schmachtenberg, R., Hörner, M., Jerez-Longres, C., Mohsenin, H., Wagner, H. J., & Weber, W. (2021). Synthetic biology as driver for the biologization of materials sciences. Materials today. Bio, 11, 100115. https://doi.org/10.1016/j.mtbio.2021.100115
Banner, A., Toogood, H. S., & Scrutton, N. S. (2021). Consolidated Bioprocessing: Synthetic Biology Routes to Fuels and Fine Chemicals. Microorganisms, 9(5), 1079. https://doi.org/10.3390/microorganisms9051079
Moore, J. C., Ramos, I., & Van Dien, S. (2022). Practical genetic control strategies for industrial bioprocesses. Journal of industrial microbiology & biotechnology, 49(2), kuab088. https://doi.org/10.1093/jimb/kuab088
Jefferson, C., Lentzos, F., & Marris, C. (2014). Synthetic biology and biosecurity: challenging the "myths". Frontiers in public health, 2, 115. https://doi.org/10.3389/fpubh.2014.00115
3. The team designed an intensive course on synthetic biology, covering its principles, tools, and applications in detail. It was delivered through different social media platforms to reach a wide audience. Later, the course was reviewed with the scientific research department at the university, and the team even suggested integrating it into the curriculum for undergraduate students.
Build a timeline of your education activities.
Divide it into two parallel tracks:
Track A: Project-related science topics (e.g., synthetic biology, genetics, disease mechanisms).
Track B: Soft Skills and awareness (e.g., communication, health literacy, sustainability).
For each age group, include three interactive entries:
Age/Stage: materials and activities delivered.
Science/ Soft Skills Event.
Project/Health Event.
End with policymakers/experts: share your science, and record the guidelines or advice you receive back.
Create a short timeline showing how each education stage influenced your:
Idea or design decisions.
Communication strategies.
Human Practices approach.
Keep it brief: 1–2 sentences per stage.
Dedicate activities to groups most affected by your problem (e.g., high-risk patients, key users, or vulnerable communities).
Collect direct quotes, questions, or reflections from participants.
Present them as a "mirror" of your journey, showing real impact and community voice.
Inquiry-Based Learning: every activity links to a clear outcome.
Kolb's Learning Cycle: move from experience → reflection → application.
Community-Based Research: value feedback as much as teaching.
By following this tool, any iGEM team can turn education into more than reporting. It becomes a living framework to track progress, reflect on impact, and design human-centered solutions.
For every event, document it using five core elements:
Short summary of the event.
Include who the audience was, what the main topic was, and the format of the session.
Explain the purpose of this event.
Link it to your project, community need, or global challenges (e.g., SDGs, public health).
Describe what actually happened during the visit/session.
Mention tools used (posters, role-play, songs, workshops, demos, etc.).
Keep it factual and concise.
What you learned from the event.
How it changed your approach, communication style, or shaped your project design.
Skills gained (e.g., public speaking, adapting science to audience level).
What participants gained in knowledge, skills, or awareness.
Include quotes, feedback, or reflections if possible.
Add a pre/post graph to visualize the effect of the event on their knowledge.
Makes every event structured and comparable across the project.
Forces teams to reflect on both sides: what we gave and what we got back.
Captures both quantitative (graphs, scores) and qualitative (stories, quotes) impact.
Grounded in trusted frameworks:
Kolb's Experiential Learning Cycle → experience → reflection → application.
Inquiry-Based Learning → questions and curiosity drive deeper understanding.
Community-Based Participatory Research (CBPR)
Education Evaluation Models (input–process–outcome–impact) → make results measurable and clear.
This approach turned our education page into more than a record. It became a way to show impact, share lessons, and build trust — a method any future iGEM team can rely on.
To support future iGEM teams in building strong entrepreneurship documentations, we created a structured tool that documents the steps we used for PRESS. This tool transforms our work into a replicable framework that other teams can adapt to their own projects.
We have created an SDG presentation template that other iGEM teams can use. This template provides a simple and consistent way to showcase their SDGs work. It is organized into four parts: Main Goal, Long-Term Impact, Positive and Negative Interactions, and Stakeholder Feedback.
Main Idea
The direct connection between PRESS. Present statistics that highlight the problem's significance and the role of PRESS in solving this problem.
Long Term Impact
We consider the future outcomes of PRESS across three key aspects:
Postive and Negative Interactions with other SDGs
How our work achieving a special goal can contribute to the progress of other SDGs, identifying its potential advantages and drawbacks on other SDGs.
Stakeholder Feedback
Helped us to refine our approach to be more sustainable for our world and impact more SDGs.
Finally, after we have shared this style with iGEM UM-Macau team. They highlighted how this style made it easier to understand the value of PRESS in relation to SDGs. Their positive feedback confirmed that our templatesuccessfully delivers both depth and simplicity.
We developed an Asthma-Friendly City Guidebook which is designed to provide practical recommendations on how to design sustainable cities that not only protect the environment but also safeguard human respiratory health, reducing the risk of asthma and other related conditions.
To make SDGs more engaging, we created an SDGs Magazine called “SDGs Daily Tips”. This magazine uses simple illustrations and powerful short messages to show how small daily actions can create a lasting impact on our future.
We also created a Podcast Series to explore SDG challenges through storytelling. We have created five episodes, each one highlights a different theme showing how our project addresses sustainability issues. In this series, PRESS is portrayed as a hero who takes on a specific SDG challenge and solves it.
The AFCM-Egypt iGEM 2025 team developed an innovative Lactobacillus plantarum-based platform for RNA therapy targeting asthma, extensible to other diseases. We contributed novel parts (MV-RNA Loader, co-BERA), a safer Listeriolysin O variant, and a biosafety system with kill switches. Our freeze-drying protocol ensures probiotic viability, while our molecular analysis platform, with risk assessment and machine learning, aids safe protein evaluation.
An optimized inhaler and PULMORA modeling framework support therapeutic delivery. Educational efforts, including songs, games, and documentation tools, enhance community engagement. These contributions provide adaptable tools for future iGEM teams, advancing safe synthetic biology.
