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Safety

A future in which progRAM and other methods of storing data at the molecular level are used on a daily basis opens the door to many improvements in people’s lives. However, such a method poses various security challenges that our team has addressed. Although the project itself is still highly foundational, anticipating problems and potential threats at this early stage provides a clear vision for the future and shows the responsible way that every scientist should consider.

Safety in the Lab

Our team shares iGEM values and high standards for biosafety and security, and our project is in full compliance with all relevant safety rules and policies of the iGEM competition, our university, and other authorities.

The team conducted its work in the laboratories of Prof. Dr. Gil Westmeyer at the Munich Institute of Biomedical Engineering of the Technical University of Munich. We received general laboratory safety training and specialized training in working within the biosafety level 1 (S1) environment, particularly in relation to bacterial and mammalian cell cultures, as well as laboratory equipment training. All experiments were carried out according to established standard operating procedures, and only after thorough communication with our supervisors and instructors. During our work on the project, we exclusively performed experiments with approved cell lines and organisms, including E. coli DH5-alpha strain and mammalian HEK293T cells. All biological materials and hazardous chemicals were handled in designated areas with the necessary safety equipment, such as biosafety cabinets and chemical fume hoods. Most of the chemicals used in our experiments did not require special safety measures, except for standard hazardous materials typical to molecular biology, such as DNA stains, antibiotics, magnetic beads, and highly concentrated acids and bases. We followed proper decontamination procedures for all biological waste by autoclaving contaminated materials and disposing of chemical waste according to the provided instructions.

Additionally, we implemented risk management strategies to mitigate potential human error, strictly adhering to lone worker policies, after-hours regulations, the use of personal protective equipment, and enforcing physical access controls and medical reporting.

We are well-informed about and compliant with all laboratory regulations in Germany and at TUM, particularly the guidelines outlined in BGI 850-0, §14 of The Hazardous Substances Ordinance (GefStoffV). We are also familiar with the emergency and safety contacts within our institution. In any situation where our project posed potential hazards or risks, we would promptly consult with our laboratory safety officer, Josef Hintermair.

Project-based Concerns

The initial goal of our project is to provide an RNA-based molecular recording system in contrast to currently available DNA-based alternatives. Our system, ProgRAM, would thus allow for transient and less disruptive in vivo molecular recording and, in principle, already offers several safety benefits over the state of the art.

Nevertheless, as part of assessing potential risks associated with (mis)use of our project, our team carefully evaluated several concerns. While developing the progRAM system, we focused on the challenges that could arise from using an RNA-editing system.

Due to our project utilizing the deamination capabilities of the dCas13b-ADAR2 RNA-base editor system, we considered the potential risk of unintended modifications to the cell’s genetic material, particularly its transcriptome. These concerns were shared by the wider public, as reflected in the results of an open survey conducted to gauge public opinion on molecular recording. You can find more information about the survey and public concerns on our Human Practices page.

To evaluate these concerns, we consulted the relevant literature and its primary authors to accurately assess the extent of off-target effects as well as strategies to mitigate them as highlighted on our Human Practices page. These strategies include, for instance, using less potent deamination machinery (REPAIR v2 with crucial modifications within the ADAR2 catalytic domain; Cox et al., 2017), designing parts with low sequence identity to endogenous RNA molecules as explained on our Modelling page, and exploring the possibilities of induced Cas13b-ADAR2 expression (inducible promoter) or assembly (split Cas13b systems), which is highlighted on our Project description page.

A more detailed description of the safety aspects of our project can also be found in the Safety Form.

References

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