Project

In France, research in synthetic biology and genetically modified organisms is regulated by the legal framework of the European Union. According to the Directive 2001/18/EC¹ and Regulation 1829² from the European Parliament, the use of GMO must be classified and carried out following an authorisation. In the Directive 2009/41/EC³, the European Union defines the “contained use” GMOs as “any activity in which micro-organisms are genetically modified or in which such genetically modified microorganisms (GMMs) are cultured, stored, transported, destroyed, disposed of or used in any other way, and for which specific containment measures are used to limit their contact with, and to provide a high level of safety for the general population and the environment”.

To achieve proof-of-concept for our project, ExoSwitch, we used genetically modified Escherichia coli to express toehold switches and human miRNAs, which correspond to the “contained use” category. All the experiments including genetically engineered E. coli were safely performed in a class 1 laboratory (BSL-1). In addition, we chose E. coli as a chassis, because the strains we manipulated (NEB10-beta and BL21(DE3)) are non-pathogenic. Our toehold switches can not be expressed without T7 RNA polymerase, which is rare and is encoded by plasmids with kanamycin or ampicillin resistance cassettes. All manipulations were performed with appropriate Personal Protective Equipment (PPE)⁴, including but not limited to gloves, lab coats and safety goggles. Bacterial manipulation was performed under a ventilation hood dedicated to bacteria manipulation.

Additionally, we performed exosome extraction experiments to verify the presence of our target miRNA in target cancer cell culture. For that, we cultivated human cancerous cells (HeLa and MCF-7 cell lines). Both cell lines are non pathogenic for humans. Still, any work with these cells was performed in a laminar flow cabinet in a biosafety level 2 laboratory (BSL-2) to prevent potential health risks for our experimenters and other colleagues in the lab. No other usage of those cells were planned beyond exosome extraction. In addition to the mandatory use of standard PPE mentioned above⁴, any experiments in the BSL-2 were performed under the close supervision of at least one supervisor. Since we manipulated genetically modified microorganisms (GMM) and cell-lines we properly dispose of them in the biological waste bin, and DASRI (déchets d'activités de soins à risques infectieux) bins⁵. All the bins were disposed of by a contractor to be destroyed. Additionally, we were informed where to find qualified first aid responders in case of an emergency.

Lab

Our wet lab team is composed of master students, who have received appropriate laboratory and safety training during the relevant University courses and classes. All team members received additional safety training in the lab where we performed our experiments.

All experiments have been conducted in the T3S lab (Environmental Toxicity, Therapeutic Targets, Cellular Signaling and Biomarkers) supported by the Inserm (French National Institute of Health and Medical Research), the CNRS (French National Centre for Scientific Research) and Université de Paris. Our team was under constant supervision of senior researchers, who ensured compliance with good safety and laboratory practices, as they have extensive expertise in the relevant areas of experimental biology. The supervisors were always informed about the work that we were carrying out.

Every lab team member utilised a personal lab coat, wore gloves and safety goggles while doing any experiments. In respect of the outside world, the waste was managed carefully. All biological materials are carefully placed in yellow "DASRI" (Infectious Medical Waste Activities) bins⁵ specific for biological matters, and then safely treated and disposed of by Université de Paris.

All of this put together illustrates how the laboratory security and safety measures are seriously taken into consideration by the UParis_BME 2021 team.

Human practice

For the Integrated Human Practices, we needed to ensure the security of our data, and whether it is collected in an appropriate manner. We mainly relied on the online surveys (via Instagram and Google forms) to gather insights from our target populations. Firstly, we chose to anonymize our survey to protect participants’ identity and other personal information. We used the data to gather statistics and use participant answers to learn what topics we should cover on our social media, and what interests our key audience. We carefully informed the participants participating in the surveys that we’re using their data in order to find an answer or highlight some key points or surprising results we could find. According to the European General Data Protection Regulation (GDPR)⁶, we offered the participants an option to request modification and removal of their data using an associated tag, unlinked to the person’s identity.

Covid-19

In the context of the Covid-19 pandemic, our team carefully respected the rules of social distancing and personal hygiene⁷. All of the team members had to fill in official documents to be allowed to come on campus and perform the required activities for our project. First and foremost, we minimized the number of people coming to the campus, avoiding unnecessary social gatherings. Whenever we had to be present at the University, we worked in a ventilated room wearing facemasks and maintaining the 6 feet distance whenever possible.

All of our members are fully vaccinated. Additionally, we all utilise the TousAntiCovid app on our phones⁸. This application was launched by the French Government to facilitate the timely notification of people who have been in contact with a person tested positive to Covid-19, and to accelerate their self-isolation and medical care.

In addition, if one of our members felt any COVID-19symptoms, he/she would not come to the face-to-face meetings or to the laboratory and perform an antigen test before returning to the campus. A covid-19 testing centre was open on site throughout the summer.