Team:BOKU-Vienna/Safety

Team:BOKU Vienna - 2020igem.org


Safety first!

Always having safety as a top priority is crucial in research projects. One besides many responsibilities of scientists is to work safely and put themselves, the community and the environment to the least possible amount of risk.

Always having safety as a top priority is crucial in research projects. One besides many responsibilities of scientists is to work safely and put themselves, the community and the environment to the least possible amount of risk. For this, risk evaluation prior to the project as well as continuous risk assessment throughout the whole project needs to be done. We gave our best to put safety first throughout the project, designing our project with considerations to safety aspects and also putting a focus on safety issues related to our project. We were able to learn a lot about which aspects need to be considered working with GMOs and when thinking about implementing our project in the future.

Safety instructions prior lab work

All team members who worked in the laboratory received safety instructions by our supervisor Dipl.-Biol. Dr.sc. ETH Zürich Martin Altvater. Besides general safety information he also gave us a laboratory tour and showed us all the equipment. Only team members who received this safety training and signed a safety form afterwards received an entrance card for the lab. Additionally, the students who worked in the lab of course have strong knowledge on how to work safely in the lab through several laboratory courses throughout their studies. For critical equipment we got additional instructions from our supervisor and they then were only used by students who received this special training.

Safety measures in the lab

Of course, we applied high safety standards in our laboratory. Besides wearing lab coats, gloves and safety glasses at all-time while working in the lab, additional protective equipment was used: glasses with a UV filter for working with the UV chamber, protective gloves for working with the autoclaves, heated materials or when using the -80°C freezer.

In order to ensure that our work was as safe as possible we cautiously evaluated our used chemicals before we actually started our laboratory work. This was done by checking and comparing the respective safety data sheets. As an example, we used SYBR safe and peqgreen as DNA stains for electrophoresis rather than the toxic compound ethidium bromide. More information can be found in our Safety Form .

Despite trying to avoid potentially harmful substances there were still some toxic chemicals which we had to use. These substances (like imidazole, chloroform and ethanol) were stored separately in a poison cabinet.

We also had a separate lab area where we conducted our gel electrophoresis with the DNA staining dyes peqGreen and SYBRSafe.

Additional safety measures due to COVID-19

Due to the special situation with COVID-19 we took additional measures in order to act responsibly and minimize the risk to contribute to the spread of the virus. Despite wearing face masks and using disinfectants for hands regularly, only a limited number of students were allowed to work in the lab at the same time which ensured having an appropriate safety distance. Also, we had an attendance list where everyone who was in the lab had to register. By this, we made sure to know who has to be quarantined in case of a COVID-19 infection of a team member.

Use of safe organisms

When we chose the organisms we wanted to work with within our project it was important to us to reduce potential risks to a minimum. Thus, we decided to carry out all our experiments with organisms classified to Risk Group 1. (E. coli DH10b, BL21(DE3), bacteriophage T7) Organisms of this risk group normally only have restricted risks and pose only a minimum threat to the wellbeing of people working with them, as well as on the community and the environment when released.

Of course, we do not release any modified organisms into the environment. However, if it should happen accidentally these organisms would cause less threat compared to organisms of other risk groups. Also all organisms we used are well established within our department. Since all these organisms were also listed on the White List of iGEM there was no Check-in required for the competition.

Safety considerations in project design

The integration of a safety mechanism in our project design seemed to be a necessity since we aim for our GMO to be applied in the human body in the future. With the help of an appropriate safety mechanism we hope to not only ensure control over the given dose in the treatment of patients but also increase acceptance within the population.

Our solution: We produce a phage which is not able to self-replicate: We replace the gene coding for the major capsid protein, essential for phage assembly and thus replication, with human plasma gelsolin. This is done by using the lambda RED system (you can find more details about this on our project description site).

Without the ability to replicate no viable phages can be produced outside of our production strain. The production strain itself carries the gene for the major capsid protein on a separate plasmid and by this is able to generate viable phages.
This mechanism is also highly improbable to fail because our transgene for gelsolin is unlikely to mutate into the major capsid protein or an equivalent protein within one generation. Beyond that, this safety control can easily be transferred to other kinds of phages and makes a potential admission as a therapy more likely.