Introduction
Safety is not an experiment for iGEM Thessaloniki. It is a commitment.
Safety in a laboratory setting is of paramount importance because it includes procedures and policies designed to protect scientists' health , avoid accidents, and preserve the validity of experiments and research. Thus, safety was a priority during all our lab procedures.
Considerations and extra safety measures
Placing the safety of others and ourselves at the forefront of our iGEM journey, we began with comprehensive training and education in order to become well versed in the lab's safety procedures, emergency protocols, and the proper handling of equipment and hazardous reagents, including ethidium bromide, acrylamide, etc.
During our experiments in the lab environment, each and every member was required to wear gloves, lab coats, and safety goggles. The workspaces were well ventilated, sterilized, and equipped with the necessary safety equipment. The commitment of everyone to abide by the safety standards and to foster a culture of awareness was the cornerstone of our lab safety.
In particular, we were extremely careful to follow the sterilization protocols during experiments with naked oligonucleotides because direct contact with naked DNA or RNA may activate the human immune system or cause mutations in the infected cells. Moreover, by adhering to safety protocols during bacterial transformations, we minimized the risk of accidental exposure to harmful substances, contamination of the work environment, and the spread of genetically modified organisms. It is also worth mentioning that the specific microorganisms we used belong to risk group 1, which are unlikely to cause harm to the environment or health problems for humans.
Pottential toxicity to human from the microorganisms we worked with
Bacillus subtilis
However, we did extensive research about the toxicity of the microorganisms. In particular, the toxin BsrG, a 38 amino acid protein, is part of a type I toxin-antitoxin system (TA) in Bacillus subtilis. It can cause cell lysis and programmed death. BsrG is mainly found in prophage elements and has been shown to be toxic when overexpressed. The biological role of TA is not yet clear, but it may be involved in maintaining genetic elements, adapting growth to environmental stress, or forming persister cells. BsrG gene expression can cause morphological abnormalities, nucleoid condensation, and global inhibition of transcription and translation. It also stimulates fatty acid biosynthesis, leading to cell lysis. The direct cellular target of BsrG is not yet identified. The toxin BsrG is classified as GRAS (Generally Regarded As Safe) in the laboratory, but its handling is considered with extreme care.
Cyanobacteria
As far as the cyanobacteria,
Biosafety seminar and Safety training
Additionally, our principal investigator, co-principal investigator, instructors, and advisors served as safety guides, supervising our experiments to prevent accidents. Furthermore, the biosafety officer of our institution, Prof. Dimitra Lambropoulou and the managers of the Integrated Environmental Management program at the Aristotle University of Thessaloniki were always willing to provide safety instructions and guidance.
In particular, Prof. Lambropoulou was so interested in delving into the biosafety world that she organized a training course for us. During our training, we discussed the crucial topic of chemical and reagent disposal within our laboratory in great detail. We stressed the critical role that ethical waste management plays in reducing dangers to the environment and human health. The thorough labeling of hazardous products, the proper segregation of waste into appropriate containers, and the safe sealing and marking of these containers to minimize inadvertent exposure were all topics of detailed talks.
Managing the risks of our project
Throughout our iGEM journey, a fundamental aspect of our project has been ensuring that it is environmentally friendly and does not harm the local ecosystems. Thus, our safety valves included our hydrogel biopolymer being biodegradable and having antimicrobial properties, while the microspheres’ consortium of genetically modified bacteria is to be killed before the natural degradation of the hydrogel, guaranteeing the safety of the ecosystem.
Is our hydrogel harmful to the environment?
We used chitosan and pectin as polymeric blocks in the hydrogel matrix. Chitosan is derived from chitin, a natural biopolymer found in the shells of crustaceans, like shrimps, and pectin is a complex carbohydrate present in the cell walls of fruits, especially citrus fruits. They are biodegradable and non-toxic alternatives to synthetic chemicals. Additionally, the synthetic peptide we used for the crosslinking of the hydrogel will eventually degrade due to its exposure to various environmental factors.
Is our bacterial consortium harmful to the environment?
As far as the bacterial consortium is concerned, we used two kill switches to ensure that the introduction of the genetically modified organisms into the environment, a practice that raises concerns primarily due to potential ecological and human health risks, is avoided. When IPTG is depleted from the nutrient medium, the induction of toxin production is triggered.(You can read more about it in our Design page) Additionally, IPTG is not commonly found in the natural environment, especially in the area around the nanoparticles in the hydrogel matrix. All the above factors, synergized with the incorporation of the antibacterial peptide elucidate that the release of bacteria into the ecosystem will undoubtedly be avoided, given their inevitable death within the hydrogel. We really hope that all this work has had an impact on students and other individuals interested in participating in scientific research, causing them to actively seek information on and abide by the most recent biosafety regulations.
Animal repellent
Animals are critical for the use of our biopolymer. Animals may devour our hydrogel, destroying our efforts to restore the burned-out area or even damaging themselves. We offer the concept of a well-known agricultural animal repellant, which is a device that assists farmers in defending their crops from animals by using a servo motor operated by an ultrasonic sensor. When the ultrasonic device detects an animal, it sends the information to Arduino. Arduino enables deterrent circuits that generate varying kHz frequencies to repel animals. The kHz frequencies are not harmful to humans.
Biosafety and Outreach
During our conference “In the World of Synthetic Biology”, one of our main goals was to underscore the importance of biosafety and biosecurity. Thus, the speakers presented the importance of biosafety during lab procedures, and during our workshops, we discussed with the audience specific biosafety measures for our own experiments. This way, we also addressed ethical concerns related to biosafety, such as potential environmental consequences. Moreover, we prepared a questionnaire in order to ascertain the level of knowledge in biosafety of our peers. We would also like to note that we informed the general public about biosafety issues at most of our other Human Practices events because one of our main goals was to highlight the importance of biosafety, ensuring that future researchers grasp its significance in safeguarding both scientific progress and public health.
