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Team UCopenhagen 2024: Parts Collection of MilkClear

Introduction

Our parts collection is a comprehensive and innovative toolkit designed to address critical environmental and health challenges, specifically focusing on the detection and detoxification of harmful contaminants such as polycyclic aromatic hydrocarbons (PAHs), dioxins, polychlorinated biphenyls (PCBs), and bisphenol A (BPA) . Leveraging synthetic biology, our collection includes an array of receptor modules, reporter modules, and detoxification enzymes, enabling the development of highly specialized biosensor devices. These devices not only detect contaminants but also actively detoxify them, making our parts both practical and forward-thinking solutions to address contamination in breast milk.

The importance of developing a biosensor that not only detects but also actively detoxifies contaminants in breast milk cannot be overstated. While detection is a crucial first step in identifying harmful substances, it only highlights the problem without offering a direct solution. By integrating a detoxification system, we take this innovation a step further, enabling immediate action to neutralize and remove dangerous compounds such as bisphenol A (BPA), polycyclic aromatic hydrocarbons (PAHs), and dioxins. These contaminants are known for their endocrine-disrupting effects, which pose serious health risks to infants, who are particularly vulnerable during their developmental stages. Our dual-function device called SUPERMOM not only provides mothers with peace of mind by ensuring that harmful substances are identified but also guarantees a practical and efficient solution to detoxify the milk before it reaches the baby, ensuring that the nutritional and health benefits of breastfeeding are preserved. This combination of detection and detoxification offers a comprehensive approach to safeguarding infant health in an increasingly contaminated world, setting a new standard in maternal and infant care.

Our contribution to the iGEM community

We have designed and submitted a collection of modular, well-characterized parts to the iGEM community, significantly expanding the toolkit for environmental sensing and detoxification. These parts are fully compatible with existing iGEM standards, enabling teams to incorporate them into a wide range of projects. Below are the key ways we contribute to the iGEM community:

1. Biosensor Modules : Our parts include receptor modules, such as pRET2-LexA-ERα(LBD), which allow teams to detect endocrine-disrupting chemicals like BPA. These receptor modules are engineered to bind to specific contaminants and initiate a measurable reporter response, using constructs like Lex6Op-pLEU2-NanoLuc to output luminescence when contaminants are present. This system provides a ready-to-use platform for teams aiming to detect environmental pollutants.

2. Detoxification Systems: In addition to detection, we offer detoxification modules, such as CYP1A1-pGAL1/10-POR and UGT2B15, which metabolize and neutralize harmful compounds like BPA, PAHs, and dioxins. These parts are especially useful for projects aiming to create solutions that go beyond detection to actively remove toxins from the environment.

3. Modularity and Flexibility: All parts in our collection are designed with modularity in mind, enabling teams to mix and match receptor modules, reporter modules, and detoxification enzymes to create customized biosensor systems. This flexibility makes our collection an essential resource for any iGEM team working on environmental sensing, bioremediation, or public health projects.

4. Cross-Project Usability: Our parts collection is adaptable to various chassis and biological systems, from Saccharomyces cerevisiae to other eukaryotic and prokaryotic hosts. This cross-compatibility ensures that iGEM teams from different domains can easily integrate our parts into their work, providing a universal toolset for detecting and managing environmental contaminants.

Showcasing the Impact of our Parts Collection

By contributing with this collection of receptor, reporter, and detoxification modules, we provide the iGEM community with tools that not only enhance current biosensing capabilities but also empower future teams to build more advanced, multifunctional systems. The potential applications of our parts extend from environmental monitoring to public health, making this collection a significant step forward in the fight against pollution and harmful chemicals. Our work encourages the iGEM community to explore synthetic biology as a practical solution for real-world environmental challenges, ensuring the well-being of future generations.

Devices

The Devices Table outlines the different biosensor devices developed for the detection and detoxification of various environmental contaminants. Each device incorporates specific receptor modules and reporter modules, tailored to detect particular compounds like polycyclic aromatic hydrocarbons (PAHs), dioxins, polychlorinated biphenyls (PCBs), and bisphenol A (BPA). For instance, devices BBa_K5477041 and BBa_K5477042 are biosensors designed to detect PAHs, dioxins, or dioxin-like PCBs, while devices BBa_K5477043 to BBa_K5477045 are specialized for BPA detection. Additionally, BBa_K5477046 introduces the SUPERMOM device, which integrates both detection and detoxification modules to simultaneously identify and neutralize BPA in breast milk. The final device, BBa_K5477047, is a detoxification device targeting PAHs, dioxins, and PCBs through the use of CYP1A1 and UGT enzymes, which metabolize these contaminants into safer, water-soluble compounds. This array of devices demonstrates the versatility of the biosensor platform in addressing diverse environmental and health-related issues.

Results

Composites

The Composites Table showcases the various composite parts that make up the functional modules of each biosensor device. Each composite part consists of a combination of promoters, receptors, or detoxification modules designed for specific detection or detoxification tasks. For example, BBa_K5477023 and BBa_K5477024 feature receptor modules tailored to detect PAHs, dioxins, and dioxin-like PCBs, while BBa_K5477027 and BBa_K5477028 are optimized for BPA detection through engineered estrogen receptor modules. Additionally, the composite parts BBa_K5477035 to BBa_K5477039 focus on detoxification, containing enzymes such as UGT2B15, UGT1A1, and CYP1A1 that metabolize harmful chemicals into less toxic or more easily excretable forms. These composite parts serve as the building blocks of the devices, providing the necessary functionality to detect and detoxify environmental contaminants.

Results

Parts

The Parts Table lists individual genetic components, including promoters, coding sequences (CDS), and regulatory elements, that make up the biosensor and detoxification systems. Promoters such as pRET2 and pSTE12 are used to drive constitutive or inducible expression of the receptor and detoxification modules, ensuring that the biosensors are responsive under the desired conditions. The table also includes various coding sequences for receptors like AhR, ARNT, and LexA-ERα, which are central to detecting endocrine disruptors like BPA and other toxic chemicals. Detoxification enzymes such as CYP1A1, UGT2B15, and CYP3A4 are also featured, responsible for metabolizing and neutralizing harmful substances. These individual parts allow the biosensors to be highly modular, enabling customization for different contaminants while maintaining efficiency and compatibility with standard assembly methods.

Results