SAFETY

Our main organism, a genetically engineered yeast, is designed to detect and respond to aflatoxins. Through a receptor activated by ligand-induced dimerization, our yeast triggers a response involving the production of a reporter protein and the overexpression of high-affinity nanobodies on its surface, which adsorb and remove the toxin from various environments. This enables rapid aflatoxin detection and enhanced adsorption capacity for both food and in vivo applications.

We strictly followed iGEM's rules and policies. We did not do any prohibited activities, such as using organisms from Risk Group 3 or 4, releasing or deploying genetically modified organisms outside the lab, or testing our product on humans (including ourselves). All of our work is covered by iGEM's White List, which includes organisms, parts, and activities. Additionally, we did not conduct any work that required advanced permission from iGEM. Our project involved laboratory work, conducted in a biosafety level 2 laboratory. In order to handle biological materials, we used an open bench, a biosafety cabinet, and a chemical fume hood, following strict safety protocols documented with photos to ensure compliance.

We worked exclusively with Escherichia coli NEB10β and Saccharomyces cerevisiae GM01, both of which are low-risk organisms. All parts used in our project, including the DNA ordered from IDT (member of the IGSC), are safe and pose no hazards. We performed molecular cloning in Escherichia coli and later implemented the constructs in Saccharomyces cerevisiae for assays. These assays included receptor functionality testing and further experiments for aflatoxin detection and adsorption. To construct our receptor, we selected domains from the Mus musculus erythropoietin receptor (EpoR). To enable polycistronic expression in S. cerevisiae, IGG6 will be used, an optimization of IGG1 from Glarea lozoyensis. Nb28-S102D, an improvement of Nb28 VHH produced in Vicugna pacos, were used for capture of aflatoxin. In addition, we utilized chemicals such as aflatoxin B1, mutagenic agents, and corrosive substances under controlled conditions to minimize risks.

Our supervisors, in collaboration with the Biosafety Guidance Service at the University of Málaga, provided extensive training on good microbial practices and the handling of hazardous substances like aflatoxins. All team members received comprehensive training on lab safety, including rules for lab access, biosafety levels, microbial techniques, emergency procedures, physical and data security, and dual-use concerns. We also implemented an accident reporting system, inventory controls, and lab access restrictions. Additionally, we received project-specific safety training, particularly focused on handling aflatoxin B1 safely. This included personal protective measures, proper labeling, and disposal protocols for hazardous materials. We comply with the safety regulations established by the University of Málaga and relevant national and international guidelines.

Data Management

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The data we collected from people complies with applicable regulations to ensure privacy and confidentiality protection. The emails were not collected and we guarantee the absolute anonymity and secrecy of the answers in strict compliance with the Organic Law 3/2018 of December 5, 2018, on the Protection of Personal Data and Guarantee of Digital Rights (LOPD) and the provisions of Regulation (EU) 2016/679 of the European Parliament and of the Council of 27 April 2016 (RGPD) on the protection of Personal Data.

Project risks

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Handling substances like aflatoxin B1 poses potential risks to team members. Also, the knowledge or the final product generated in this project could potentially be misused, for example, by concentrating aflatoxins for malicious purposes.

To minimize the risks associated with our project, we have implemented the following preventive measures:

• We strictly followed Harvard University's guidelines for aflatoxin handling, along with safety protocols from Sigma Aldrich.
• We only used aflatoxins in the final stages of our project. In vivo application would require preliminary studies in animal models to ensure there is no adverse effect on natural microbiota. We comply with GMO regulations in Spain (Law 9/2003 and Law 14/2011), which requires approval from the Ministry of Science for the restricted use and voluntary release of GMOs.
• We used a strain of Saccharomyces cerevisiae with multiple auxotrophic requirements, preventing its growth and spread outside the lab. This strain only survives in the presence of essential nutrients that are not available in the environment. Our lab also has strict protocols for the disposal of GMOs and aflatoxins: all cultures and materials in contact with aflatoxins are sterilized using bleach and autoclaved for safe disposal, following the WHO Laboratory Biosafety Manual and New England Biolabs guidelines for handling E. coli. We carefully labeled and tracked all materials used.
• All team members used appropriate PPE, including lab coats, gloves, and protective eyewear, particularly when working with hazardous chemicals like aflatoxin B1. Any spills involving aflatoxins or other hazardous substances are immediately processed, discarding or, if not possible, cleaning very consciously. We evaluated the potential misuse of our technology and consulted with experts how to mitigate these risks. We ensure that our yeast’s modifications do not result in harmful consequences by restricting its application to laboratory environments.