Team:KU Leuven/Safety
BLADEN
Safety
Introduction
Safety forms an important aspect of all team projects. To ensure the smooth sailing of our project and the health and safety of all members and personnel involved, we have taken many steps to secure their safety as you can read below. Generally, we have analyzed the risks associated with our laboratory experiments, followed general and lab-specific safety guidelines. We have also tried to analyze the various ethical and biosafety concerns of our project. All lab experiments and meetings were held in accordance with Covid Safety guidelines as laid down by our institution, KU Leuven, and the Belgian Government.
Lab Safety
The members of our wet lab team worked in Professor Vitor Pinheiro’s lab at the Rega Institute for Medicinal Research, and Professor Filip Rolland’s Laboratory of Molecular Plant Biology. All team members involved in laboratory experiments underwent a mandatory Health and Safety training session. Risk awareness, health surveillance, purchasing chemical products, and the first point of contact for First Intervention and First Aid, were discussed. Emergency procedures in case of a fire or exposure to chemicals, location of fire extinguishers, and the emergency phone number were made aware to all members of the lab subgroup.
Experiments were conducted within standard lab hours, barring exceptional cases where they had to be conducted outside these hours. In such cases, members worked in sub-teams of two and followed the required lab protocols to ensure safety while working. Wearing appropriate personal protective equipment such as lab coats, gloves, and masks was strictly followed. Training to use the chemical and laminar hoods was provided to all members of the lab sub-group. The storage of purchased chemical products was as per the laboratory protocols. Waste materials produced were disposed of as mandated by the particular lab. The use of specific equipment like confocal microscope, PCR machine, etc. was done as per the safety protocols mandated by the labs.
Biological Safety
The experimental risks were assessed based on the different organisms used and experiments conducted. Escherichia coli, Agrobacterium tumefaciens, and Tobacco BY-2 cells are the organisms used in our project. The lab-adapted, unmodified E. coli presents a low risk to our team members and the environment if exposed. While the Agrobacterium used has a good track record of laboratory use and presents a low risk to our team members as well as mammalian hosts in general, it is known to pose a small risk if exposed to plant cells. However, the constructs being introduced, will not increase the pathogenicity of the Agrobacterium in the environment.
The antibiotic resistance cassettes can represent a potential source of personal, community, and environmental risk if they can be mobilized via horizontal gene transfer to other locally adapted hosts. However, EvolvR components (Cas9 and error-prone DNA polymerase) are not active in the absence of a guide RNA and the guide RNAs are being engineered to target genes that are not present in mammals (GFP and glutamine synthetase). The cultured BY-2 Tobacco cells used did not present any significant risks to the researcher or the environment in general.
Design Safety
In the design and assembly of the CellES device, technical and organizational safety measures must be followed to ensure the safety of the personnel involved, and prevent any environmental pollution. Some standards that need to be adhered to include Safety of machinery – General principles for design – Risk assessment and risk reduction (ISO 12100:2010), The law voltage directive (2014/35/EU), The General Product Safety Directive (2001/95/EC). To further ensure safety, the device must be assembled with additional conditions: proper illumination, fresh air supply, removal of hazardous gases, suitable air temperature and humidity, cleanliness. Device parts and containers must be labeled, while sterilized components and hazardous materials, must be used in specialized rooms.
Biosafety and Ethical Concerns
The Nuffield Council for Bioethics has recognized four ethical concerns with Genetically Modified Organisms (GMOs): potential harm to human health, potential harm to the environment, negative impact on traditional farming, and excessive corporate dominance [1]. Our project BLADEN contains the same safety and ethical risks as GMOs but does not present any new concerns to the current landscape of issues. We believe that the possibility of generating new plant variants using CRISPR technology is relatively safe, given the well-known accuracy of this technology. The engineering of organisms can be pursued safely while minimizing the risk of obtaining undetected, undesired mutations.
Safety during Covid
Working on the iGEM project during the Covid-19 pandemic of 2020-2021 posed interesting circumstances for both the project and the team. Nevertheless, looking back, we have tried to make the best of the different scenarios all year round. In the beginning, our team met each other via meetings on Zoom. We held many brainstorm sessions this way. When the rules relaxed here in Belgium, we took our discussions outside by taking walks around the city of Leuven in small groups of four members.
From July 2021 onwards, weekly meetings were held in our iGEM office. Lab meetings were also increasingly held in person. At all times, mouth masks were worn, and closed spaces were well-ventilated. In the labs, protective gear and mouth masks were worn at all times. Booking exclusive sessions for the use of lab equipment and hoods was a common practice. Lab work was always done in subgroups of two or three members. While conductinig outreach events and human practices events, all government-mandated rules and regulations, along with KU Leuven's regulations, were followed.
More information can be found here:
Covid Measures in Belgium
Corona Measures at KU Leuven
References
[1] Weale A. Ethical arguments relevant to the use of GM crops. N Biotechnol. 2010 Nov 30;27(5):582-7. doi: 10.1016/j.nbt.2010.08.013. Epub 2010 Sep 17. PMID: 20850572.