Innovating Safety in Synthetic Biology
Our project redefines safety in synthetic biology by engineering Escherichia coli to degrade the explosive RDX in freshwater, using a closed bioreactor system. A novel biocontainment kill switch, activated by degradation byproducts (formaldehyde and nitrite), ensures no environmental release. Operating in a Biosafety Level 2 lab, we enforce strict protocols—PPE, biosafety cabinets, and chemical safety training—to manage hazardous materials like RDX, ethidium bromide, and corrosives, setting a benchmark for responsible synthetic biology.
Explore iGEM’s safety standards at the Safety Policies page.
Laboratory Safety
Our team operates in a Biosafety Level 2 laboratory equipped with open benches, biosafety cabinets, and chemical fume hoods. All members complete extensive safety training, covering lab access, microbial techniques, chemical handling, and emergency protocols. Mandatory Personal Protective Equipment (PPE)—lab coats, gloves, and safety goggles—ensures protection, while rigorous waste management and decontamination protocols maintain a secure environment.
Project Overview
We engineer Escherichia coli (strains EC02T110, EC300110 and DH5α) to detect and degrade 1,3,5-trinitro-1,3,5-triazine (RDX) in freshwater using a bioreactor. A constitutive promoter drives an mRNA with a translational riboswitch that binds RDX, enabling xplA and xplB gene expression for aerobic degradation. Byproducts (formaldehyde and nitrite) activate a biocontainment kill switch, ensuring bacterial self-destruction.
Safety and Compliance
Our project complies with iGEM’s safety policies, using non-pathogenic E. coli strains and White List parts. We avoid prohibited activities, such as using Risk Group 3 or 4 organisms, releasing GMOs, or human testing. All DNA/RNA is sourced from IGSC-compliant providers, and no activities (e.g., animal use, gene drives) require advance iGEM approval.
Risk Identification
Our work involves hazardous chemicals: RDX (explosive, possible carcinogen), ethidium bromide (mutagen), chloroform (carcinogen), formaldehyde (mutagen), nitrite (carcinogen), ethanol (flammable), NaOH, and HCl (corrosive). Identified risks include:
- RDX exposure: Potential central nervous system damage from prolonged oral exposure.
- Chemical mishandling: Burns, toxicity, or respiratory irritation from mutagens or corrosives.
- Bacterial contamination: Cross-contamination of E. coli strains affecting results.
- Physical hazards: Injuries or fires from broken glassware, hot equipment, or unattended Bunsen burners.
Mitigation includes PPE, biosafety cabinets, chemical fume hoods, strict SDS protocols, and proper waste disposal. Our bioreactor prevents environmental release, and the kill switch ensures containment.
Future Considerations
Designed for a closed bioreactor, our project prevents environmental release. Scaling to sites like Anones Lagoon requires Puerto Rico DRNA permits and compliance with NEPA, EPA, and OSHA regulations, plus institutional biosafety approvals. Consultations with Dr. Carlos Ríos Velázquez and Dr. Patricia Ortiz Bermudez ensure robust risk management.
Safety Protocol
Our comprehensive safety protocols, covering biosafety and risk mitigation, are detailed in our safety form, available below:
Our project adheres to iGEM Safety Policies and OSHA Laboratory Safety Guidance.
