SAFETY
When working with genetically modified organisms, safety is one of the most important aspects. As our project revolves around the protozoan parasite Leishmania tarentolae we always had to make sure that our work does not pose a threat for humans or the environment.
You can find our finished safety form here.
General laboratory safety
Before we started working in the wet lab, all of our team members were briefed by a qualified supervisor on general laboratory safety measures.
All of our work took place in S1 laboratories (biosafety level 1), which means that there are no risks to humans or the environment. Nevertheless, basic safety measures must be observed, such as wearing protective clothing like lab coats, safety goggles and gloves, especially when working with toxic or carcinogenic substances.
COVID-19
Just like last year, iGEM 2021 and our project were under the great influence of the current COVID-19 pandemic. To ensure the safety of all team members, supervisors, and other laboratory staff with regard to possible infections, strict hygiene rules had to be followed while working in the wet lab. Hands had to be disinfected, medical face masks had to be worn at all times, and a safe distance was maintained wherever possible.
If team members had shown symptoms that indicated possible infection with COVID-19, such as cough, fever, or sore throat, the affected person would have to stay home and test negative for COVID-19 in order to return to the laboratory. In addition, any members who had close contact with the affected person would also be tested to prevent the spread of the virus.
Leishmania tarentolae
Safety played a major role in our search for a suitable chassis organism for our project. The unicellular winged protozoan organism Leishmania tarentolae, which was already established in one of the research groups overseeing our project, is classified as a risk group one organism and white-listed for iGEM 2021.
Since L. tarentolae is a parasite that affects lizards, it was very important for us to ensure that there was no risk to humans or the environment. Genome sequencing of L. tarentolae revealed that the protozoan parasite lacks several genes associated with pathogenicity to mammals compared with other Leishmania species.
These safety aspects, combined with the advantages that L. tarentolae has in terms of human-like protein production, provide us with the perfect chassis for our project.
Escherichia coli
The Gram-negative bacterium Escherichia coli also plays a key role in our experiments. It serves as a tool for propagating DNA to be transfected into L. tarentolae. Although E. coli is the most widely used and best studied organism in biotechnology, safety plays a very important role when handling these organisms. As with all genetically modified organisms, we must ensure that E. coli does not leave the laboratory. Therefore, we sterilize all contaminated waste with ethanol, collect everything in a separate container, and autoclave the waste. All contaminated surfaces are cleaned and disinfected as soon as the work with E. colil is finished.
Project-specific safety
The goal of our project is to establish the modular cloning system in Leishmania tarentolae to make the organism more accessible for protein production. The main advantage of L. tarentolae is its glycosylation patterns, which are very similar to those of humans, making it a promising tool for therapeutic protein production.
Since our project aims only to establish a new mode of protein production in this organism, there is no need to release the protozoan into the environment. Therefore, the risk of possible uncontrolled spread in the environment is minimized if the organism is handled according to standard safety protocols.
After establishing the MoClo system, it would be possible to place the Leishmania strains in a bioreactor to maximize protein yield. This would also be the optimal choice to avoid possible unwanted contamination, as bioreactors are considered very safe.
Ethics
Biotechnology and especially genetically modified organisms are still a very sensitive topic nowadays. Since our project aims to provide a tool to produce therapeutic proteins, it is very important for us to work responsibly and educate the public about genetic engineering
To get an overview of what people think about biotechnology and genetic engineering, we conducted a survey asking people's opinions on various aspects of GMOs.
Sources
- https://www.gesetze-im-internet.de/gentg/index.html
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6657821/
- Klatt S, Simpson L, Maslov DA, Konthur Z. Leishmania tarentolae: Taxonomic classification and its application as a promising biotechnological expression host. PLoS Negl Trop Dis. 2019;13(7):e0007424. Published 2019 Jul 25. doi:10.1371/journal.pntd.0007424
- https://www.jenabioscience.com/lexsy-expression/lexsy-background/biosafety
- https://www.jenabioscience.com/images/b3e879b381/Genome_of_Ltar_NAR_2012_gkr834.pdf
- Raymond F, Boisvert S, Roy G, Ritt JF, Légaré D, Isnard A, Stanke M, Olivier M, Tremblay MJ, Papadopoulou B, Ouellette M, Corbeil J. Genome sequencing of the lizard parasite Leishmania tarentolae reveals loss of genes associated to the intracellular stage of human pathogenic species. Nucleic Acids Res. 2012 Feb;40(3):1131-47. doi: 10.1093/nar/gkr834. Epub 2011 Oct 13. PMID: 21998295; PMCID: PMC3273817.
- https://2021.igem.org/Safety/White_List
- https://www.jenabioscience.com/lexsy-expression/lexsy-background/biosafety