Safety was a top priority for our team, especially because a majority of our research took place in a biochemical laboratory setting. The lab in question was the third floor of the Molecular Plant Sciencesbuilding at Michigan State University, classified as a Biosafety Level 1 laboratory.

Once permitted to work in the lab, the team followed the safety protocol outlined in our training, including wearing long pants, closed-toe shoes, and disposable gloves. When working in a fume or laminar flow hood, team members wore safety glasses/goggles and lab coats. Additionally, safety protocols required that team members could never be in the laboratory if a graduate student, postdoctoral researcher, or faculty member was not also present. Because the lab space was shared by four different research groups, supervision was rarely an issue. On a day to day basis our research team wore nitrile lab gloves, long pants and closed toed shoes as PPE. Additional PPE of a lab coat was worn when running gel electrophoresis to protect from ethidium bromide. All bacterial work was performed in biosafety cabinets. We worked with Trichoderma spp. and Fusarium spp. in a laminar flow hood because of their high propensity for contamination of other fungal cultures.

Safe Project Design

The team made sure to use a safe, non-pathogenic chassis for our project. We transformed chloramphenicol resistance, GFP and the acds gene into our chassis Enterobacter ludwigii. This species is most closely related to Enterobacter hormaechei which is considered a risk 2 organism. We chose Enterobacter ludwigii for its lack of the acds gene, relatively quick growth, lack of innate resistance to chloramphenicol, and transformability. We decided not to include a kill-switch device in our organism because that would have required us to genetically engineer the plant it was applied to and spray a chemical onto the plants to induce endophyte cell death.

Safe Shipment

Parts were submitted in a 96 well PCR microplate, which was sealed with foil, cover placed on, placed into a Ziploc bag, and shipped in a box with adequate air cushioning. Contents were declared as non-hazardous DNA. These practices follow the iGEM part submission requirements.