TU Dresden/Safety
Safety
Laboratory ergonomics
Team member education and training was essential in order to avoid injuries in the laboratory. This involved a safety briefing from our supervisor (Dr. Frank Groß) as well as a personal walk-through of our soon-to-be work area. This meant learning everything from where the ice is kept to where isolated areas for handling ethidium bromide takes place. We have all learned proper techniques in the lab through our respective studies, internships and jobs. Therefore, we set up our equipment in such a way that multiple team members could work at the bench simultaneously.
Obtaining results in the lab meant putting in extra hours and this meant working weekends and after hours, not a foreign concept to our fellow iGEMers. In order to insure safety in the off hours, team members were not allowed to work alone and even if that meant a team member is sitting working on the wiki, no one was ever alone in the lab. In case of emergency, another set of eyes and ears was always on hand. Ergonomics is essential when setting up the lab space and thanks to great mentorship; no issues or injuries were sustained in the course of our iGEM experiment.
E. coli strains
We were given stringent guidelines as to how we should handle our E. coli strains. We used ER2738, GBO5, BL21 and K12. All of these strains are non-pathenogenic and the former pair lack common pathogenicity-related sequences and the latter have disabling mutations that prevent them from surviving in environments outside of the lab. Safety data sheets were carefully read in order develop laboratory protocols for emergencies, handling and disposing. We also used bacteriophages, which are infectious to only bacteria and in order to make sure phages were contained properly, we took proper care during the disposal of exposed bacteria.
Biobrick parts
It was important to consider the pathogenicity and toxicity of our iGEM parts we planned to submit, to provide a safety data profile of our own. All of our parts are non-pathenogenic and are non-toxic. The only concern was the HER2, which is a human protein and therefore needs extra attention as to exposure and mutated products. If we express this protein in E. coli and it underwent mutations, there are some implications that it could become harmful to the lab. Considering this protein is already implicated in a number of cancers, we took extra care to dispose of it properly once folding analysis was completed. All other bricks were from our chassis and did not have any major safety concerns, however proper care was taken to avoid any incidents in the lab.
Cleaning up our plasmids
In all of our plasmids we had antibiotic resistance of some sort in order to verify that proper conjugation occurred and to rid the population of plasmid free colonies. This was essential in all of our experiments. However, the type of resistance changed after certain plasmids were altered and once the plasmid is stable, resistance is not necessary. There still remains the issue however, of a strain caring a number of antibiotic resistances. This is a growing issue in health science and to reduce our own resistant bacteria, we inserted lox P sites outside of every antibiotic resistant gene in the plasmids we designed. Therefore once the resistance was not necessary, we could cleave it out of the bacteria and "clean up" our strains from having multiple types of antibiotic resistance or any at all. This was our way of contributing to the growing issue of antibiotic resistance bacteria in the population and hopefully provides a solution for other labs with similar concerns.