All research conducted by the iGEM team TU-Eindhoven 2023 is in accordance with the safety policies and rules from iGEM, our university, and the Dutch government. Most of the organisms and parts used and/or designed are on the iGEM 2023 White List. IL-10 was the only part not on the White list, so a check-in form was submitted before usage.

The TU-Eindhoven 2023 team has only worked in an ML-I lab and received proper training about safely working with GMOs before getting access to the lab from our university’s biological safety officer. We also received training from our lab manager about how to safely work in the ML-I lab. Furthermore, before use of any type of machine, an introductory training was done by all team members who would be using it. As personal protection, we wore lab coats, glasses, and gloves when needed. Long hear was always tied and we never wore shorts or open shoes in the lab. Waste was disposed in the appropriate bins, with chemical waste going into the black containers and biological waste into the blue containers with a yellow lid (see the Figures on the side).

For our research, we have used different E. coli strains. We used TOP10 Chemically Competent cells (NEB) for transformations of ligation products. For protein expression, we used BL21 (DE3) (ThermoFisher) cells. All the cells have been altered to be safely used in an ML-I lab. All GMO-based lab work done on bacteria was registered in the GMO notebook present in the laboratory.

We used multiple vectors, of which detailed information can be found on the Engineering Page. By fuzing different proteins and basic parts together, we were able to create many new parts that can be found on our Parts Page.

During our project, we stayed in close contact with our biological safety officer for advice on how to properly contain our bacteria during imaging experiments. We found out that we had to fix the cells before we could do any imaging experiments, since the microscopes were not in an ML-I lab. However, this was not possible for the FRAP experiments that we wanted to do. After many conversations with the biological safety officer of our lab, we eventually got the green light to do FRAP on live cells.