Team:IOANNINA/Safety
Safety in the lab and biosafety of the project were two of our main goals starting this year’s project. As a newly formed iGEM team in our University, we tried to establish the basic safety rules in all aspects of our team work for the upcoming teams in our campus. The safety steps we followed included the lab and members safety, based on the guidelines of our University and according to the legislation applied in Europe and the general biosafety underlying our project itself. Training The laboratory in which we had our experiments is classified as biosafety level (BSL) 1 and is used for undergraduate laboratory exercises. All the members of our team that worked in the lab were trained on the safety protocols followed in the laboratory, by Assistant Professor D. Alivertis, who also kindly provided us a brochure with all the safety rules established in the department. As we find that safety in the lab is a matter that should concern everyone working on a Synthetic Biology project, some of the other members of our team also joined part of this training, being introduced in the basic aspects of biosafety. The training and safety brochure included:
Lab members safety All lab members used protective gear (lab coats, protective glasses, gloves, masks) when entering the lab and dangerous materials (e.g. ethidium bromide) were used according to strict protocols. COVID-19 safety: Due to the ongoing COVID-19 pandemic, we took extra measures to ensure the safety of all lab members. We worked on small groups, used protective masks and ensured the proper ventilation of the lab. Use of harmful reagents For visualizing DNA fragments in agarose gel electrophoresis procedures we used ethidium bromide which is known to be highly mutagenic. Thus, the lab members that used this reagent were working in a fume hood, while also wearing personal protective equipment (PPE) to minimize exposure and all the proper disposing protocols were followed. Training As with every Synthetic Biology project, one of the most important issues is that of biosafety. In order for an idea to be applicable in real life, it should give the best outcome without posing safety concerns. When using a modified bacterial strain the problem of biosafety is always one of the main issues that need to be tackled. On the basis of this premise, we tried to include in our design a series of biosafety key elements, to turn this idea to an application. General microorganism information For our chassis organism we used the common non-pathogenic strain of E.coli DH5a, which belongs to the Risk Group 1 microorganisms (BSL1). We cultured two of the plasmids in the E.coli strains DB3.1 and E811 (P2 lysogen). All the strains that we handled were non-pathogenic, frequently used laboratory E.coli strains. Biosafety key features Biosafety of our proposed implemention was of outmost importance for our team during the designing process of our project. Once we had the initial idea of constructing a bacterial strain that could deactivate antibiotics present in its substrate, all of our next steps were about making this strain as safe as possible in order to have all the theoretically necessary features to make it safe before it could be tested. Our design contains three basic goals and biosafety proposals: 1. Reduce gene transfer possibility The first biosafety aspect that we took into consideration was the transfer of plasmid DNA between bacterial strains. In order to minimize this effect we decided to use a chromosomal integration protocol and integrate the gene cassettes we designed into the chromosomal DNA of the bacteria. This would minimize the possibility of horizontal gene transfer. 2. Ensuring self-destruction The second biosafety feature of our designed bacterial strain was the use of a kill switch. Kill switches are a frequently proposed solution for self-destruction of microorganisms under specific conditions. The kill switch in our project would be regulated by the presence of antibiotics, leading to the self-destruction of our strain once it has deactivated the amount of tetracyclines and macrolides in its substrate. 3. Implementation safety measure Even though these molecular mechanisms would induce the safety of the bacterial strain in the lab, we thought that for a real-life application, these two parameters would not be enough. This is why our final proposal for the implementation (See implementation) of our project supports that this strain would be used in a strictly contained environment before the stage of composting, to avoid any leakage of our microorganism in the nearby environment. 
Safety
Prologue
Lab safety
Project Safety