At the heart of our iGEM project, E-Sole I, is the commitment to safety, both within our laboratory and in the potential real-world application of our microbial foot care system. Guided by the principles of responsible research and engineering, we have implemented comprehensive safety measures to mitigate any risks associated with our work. Safety is not an afterthought; it is integral to every stage of our project, from design to implementation.
The Department of Biotechnology (DBT), under the Government of India, regulates biosafety and biosecurity for research laboratories nationwide. Comprehensive rules and regulations governing laboratory safety are accessible through the DBT's official platform, ensuring that all research institutions comply with national standards. At IISER Kolkata, we also follow an internal set of laboratory safety protocols, established and overseen by the Institutional Biosafety Committee (IBSC). These protocols are essential for maintaining a secure lab environment, reducing the risk of accidents, and ensuring the safety of all personnel involved.
Our team underwent rigorous lab safety training under the supervision of Dr. G. Lekha, a Scientific Officer at IISER Kolkata. This training emphasized the proper handling and disposal of biosafety hazards, as well as the use of essential safety equipment like biosafety cabinets, fire extinguishers, and eyewash stations. The training culminated in biosafety exams, and all team members were awarded safety training certificates upon successful completion, verifying their qualifications to conduct lab work safely. As a precaution, the laboratory was fumigated to eliminate potential airborne pathogens before starting wet-lab activities.
In compliance with Biosafety Level 1 (BSL-1) standards, all experiments related to cloning and the expression of the quorum-sensing unit were conducted in designated BSL-1 laboratories. The bacteria used in these experiments include BSL-1 strains like E. coli DH5α, TOP10, and BL21. The fungal strain used Chaetomium thermophilum, which too falls under BSL-1. For containment and risk minimization, all experiments were performed using class biosafety cabinets, ensuring that our research complies with the highest safety standards.
These stringent safety measures, along with the adherence to both DBT and IBSC regulations, ensure that all research activities are conducted in a responsible and risk-mitigated manner.
The iGEM Laboratory at IISER Kolkata is fully equipped with all the necessary safety equipment to ensure a secure and safe working environment for our research activities. The key safety features of the lab include:
Proof section details the experiments and evidence that support the feasibility and effectiveness of the design.
In the development of our microbial foot care system, E.Sole I, we have placed a strong emphasis on design safety to proactively address potential risks in both the laboratory and future real-world applications. Our goal is to ensure that the use of our genetically modified bacteria (GM bacteria) is both safe and effective for users, while minimizing risks to the environment. The following safety measures are integrated into our project design:
We initially conducted our research using the E. coli strains BL21, DH5α, and TOP 10, which are widely recognized as safe, non-pathogenic, and suitable for biosafety level 1 (BSL-1) environments. These strains were chosen for their ease of manipulation and efficiency in gene cloning, making them ideal for our lab work.
For the final product, we plan to use Lactobacillus rhamnosus as the microbial chassis embedded in the footbed. This strain is a well-known probiotic, commonly found in human gut microbiomes and widely used in health applications. Due to its non-pathogenic nature and long history of safe use, Lactobacillus rhamnosus poses minimal risk to humans, even upon direct contact with the skin. Its compatibility with the skin microbiome further ensures that it will not cause irritation or disrupt the natural microbial balance, making it an excellent choice for prolonged use in foot care.
To prevent any risks of environmental contamination or uncontrolled bacterial growth, we have designed a kill-switch (apoptosis module) that triggers bacterial cell death if the bacteria escape from the footbed or come into contact with non-target environments. This apoptosis module ensures that the bacteria do not persist outside the intended system, reducing the likelihood of unintended consequences. This safeguard is particularly important for preventing the accidental release of genetically modified organisms (GMOs) into the environment, thus ensuring both user and environmental safety.
Our containment strategy is designed to ensure that the GM bacteria remain securely housed within the footbed system. The bacteria are embedded in a nutritive biopolymer layer that is both biocompatible and non-porous, preventing their release into the external environment. The footbed structure includes a non-porous layer at the bottom to contain the bacteria and a porous layer at the top, allowing only the beneficial compound, linalool, to diffuse out. This containment system ensures that while the bacteria continue to function and produce linalool, they remain isolated from direct contact with the outside environment.
The materials used in the footbed are biocompatible and non-toxic, ensuring that they are safe for continuous contact with human skin. The porous layer allows the volatile linalool to escape while ensuring that the bacteria do not leak out. Additionally, by maintaining compatibility with the skin’s natural microbiome, our design minimizes the risk of skin irritation or infections. This makes our footbed suitable for long-term wear and safe for a broad range of users.
We understand that environmental safety is paramount in synthetic biology projects. To address this, our containment and kill-switch strategies are complemented by the use of biodegradable materials for the footbed. In the event of disposal, the materials used in the product will break down safely without releasing harmful byproducts into the environment. Furthermore, the kill-switch ensures that the bacteria will not survive outside the controlled footbed environment, further preventing any unintended ecological impact.
Ethical considerations are central to our iGEM project, E.Sole I, particularly in the field of synthetic biology. We adhere to ethical standards and comply with all regulatory requirements throughout the project.
As part of E.Sole I, we engaged with the public through interviews, surveys, and webinars, ensuring compliance with all ethical and legal standards.
From lab work to future product development, safety has been prioritized at every stage of E.Sole I.
By embedding ethical safety practices into the design, public engagement, and future production phases, E.Sole I aims to provide a safe, effective, and responsible solution for foot care.