Future Risk Considerations

In the optimal case, we would like to fully develop our project into a real product with real-world application, which would involve releasing the protein-based biosensors as a part of a small-enclosed device. If we are lucky enough to bring this forward to that stage, while all shall celebrate the success and anticipate a huge improvement in water testing efficiency and safety, many other risk considerations should also not be ignored to ensure responsible application. We have discussed the future application from the perspectives of ethical permissivity and regulatory compliance, but there are more possible associated hazards/risks to be considered and have a respective management plan devised.

Further Laboratory Experiments 1. Pathological Bacteria Testing – We may use pathological bacteria cultures, instead of only the surface proteins from less pathological strains, to make our in-lab characterisation and calibration more accurate.
2. Environmental Samples – We may need to test our products directly on the unprocessed samples collected from the real world, which would pose a high risk of contamination.
Risk Management:
To minimise the risks and alleviate the biosafety concerns in future experiments, we believe the most crucial management strategy is sufficient preparation and careful planning, thus allowing us to take any precautionary measures. At each stage, we should be doing the same things as we have done for these months of proof of concepts – thinking about and assessing all the possible risks of the procedures, parts, and organisms involved beforehand, minimising the risks by using alternative methods or materials, obtaining approval from the institutions, selecting the laboratories and equipment with the appropriate biosafety level, and practising all safety measures.

Human Health 1. Biological Hazards -- The biosensor involves the use of engineered proteins. If not handled properly, there's a potential risk of exposure to these proteins which could pose health risks to end users. During the production stage, genetically-engineered E.coli cells are also used, posing a risk to the health of production line workers upon unprotected exposure.
2. Toxicity and Allergenicity -- Any new proteins, de novo or naturally occurring, introduced into the surroundings could potentially have toxic effects on humans. It's crucial to assess the toxicity and allergenicity of the designed protein before deployment.
3. Testing Misinterpretation and False Results -- The test may have worsened accuracy and reliability during extreme storage and/or transportation condition. If the test is rendered ineffective, it may give a false negative result and mislead the end users to believe that the water tested is safe to drink. Alternatively, if the luminescent signal is not strong enough or somehow the guide is not clear, the users can misinterpret the result and be mistaken about the safety of their water. Risk Management:
We should conduct experiments to confirm that the biosensor poses no immediate hazards, toxicity or allergenicity to humans – though this is indicated by the Baker Lab paper, we should ensure that our manipulation of the sequences and the bacteriocin-derived binding motifs do not add additional risks or are incompatible with the original structures. The test kit should also be designed to minimise the chance of exposure and leakage. The reliability and stability of the test kit should also be tested under different circumstances and optimised by further polishing the molecular components and hardware components. Sensitivity and specificity should be tuned to be in a reasonable range. Finally, we should also get approved as medical devices by the official institutions.
Environment and Ecosystem 1. Ecological Hazards -- If the engineered proteins are released into the environment, unintended ecological impacts could occur. For instance, the modified proteins could interact with other organisms in ways that were not anticipated, potentially disrupting natural ecosystems. During the production stage, genetically-engineered E.coli cells are also used, posing a risk of accidental release and exposure.
2. Spread of Modified Organisms -- If the genetically-modified E.coli cells used during the production stage are not well-contained, there is a risk of them spreading beyond the intended experimental environment. This could lead to the introduction of novel traits into the environment, with potential consequences for ecosystems.
3. Waste Disposal – Considering that the biosensor kits are most likely very cheap and that the low-resource regions would have a limited capability (financial and human resources) on managing the collection of waste/used kits, the discarded kits can very easily come to direct contact with the environment. Thus, if the kits release toxic by-products when decomposing or cannot be decomposed at all, it will introduce a huge environmental problem.
Risk Management:
We should conduct experiments to confirm that the biosensor poses no immediate hazards, toxicity or allergenicity to the environment. The test kit should also be designed to minimise the chance of exposure and leakage. Safe disposal practices should be clearly written in the package of the test kit with required items (e.g. medical-level resealable bags) included.

Dual-Use 1. Misuse Potential -- Depending on the sensitivity and specificity of the biosensor, there's a possibility that it could be used for purposes other than water testing, such as detecting pathogens for harmful intent. For example, in the wrong hands, biosensors could be used to develop bioweapons capable of detecting specific individuals or sub-populations, thus enabling precision targeting and attacking.
Risk Management:
We should consult with experts in biosecurity to discuss the possibility of dual-use or misuse. The information of our design should be published with care, and the sensitive parts protected by limiting technology/information/data/physical/laboratory/stock/communication access. We should ensure monitoring along the value chain and confirm correct delivery and usage.
Society 1. Stigmatisation of Certain Regions – Regions that have high water contamination detected may be stigmatised nationally and internationally, which may hinder their economic development due to reduced private investment and population influx. As exact locations can be recorded, households with contaminated water may be socially stigmatised and bring isolation and social anxiety to the members of the families.
2. Informed Consent – The collection of data points and automatic uploading to the global database cloud may infringe on the privacy of the general public and/or the local communities, unintendedly releasing personal/community information that can be taken advantage of for ill intentions.
3. Equity and Access -- The technology should be designed and deployed in a way that ensures equitable access to its benefits and avoids exacerbating existing social or economic disparities. Policy and/or political issues in certain states/regions, or, in some cases, geographical difficulty, may cause unequal access to this water testing tool. Additionally, this unequal access to water testing can be self-reinforcing by highlighting the regions with existing data points and concentrating resources on those regions, while leaving the originally missed regions even less likely to gain access.
4. Conflicts over Water Scarcity – With the water testing tool, it is likely that less water would be identified as drinkable. If no sufficient action is taken to decontaminate the unsafe water and reduce the sources of contamination, there will be a heightened tension over more alarming water scarcity, which may lead to more violence and conflicts in the regions with contaminated water.
Risk Management:
We should consult experts in ethics and social sciences to devise a plan to minimise these risks. By cooperating with charity organisations and global institutes, we should ensure the appropriate division of available resources. Moreover, as scientists, we carry the responsibility of doing public engagement and public education on water contamination and decontamination, stimulating communication in a transparent, honest, objective, and accessible way to all different population groups. We can also collaborate with water-cleaning organisations.

Lastly, we reached out to the Safety and Security Committee of iGEM to inquire about de novo protein uses, as in our project. Taking the advice from Alonso, we ran protein BLAST of our de novo protein sequences and found no similarity to other naturally existing proteins, let alone naturally existing proteins that have the potential to pose harm. Oligo synthesis companies we have ordered the genes from also have indicated no issue with the sequences we submitted and tried to express, which indirectly reflects a relatively low possibility of risks. We have also tried to reach Baker Lab to inquire about their safety consideration and measurements on de novo proteins.

Education

We believe that, after so much work and thought putting into the relevant aspects in regard to our project, education on biosafety and biosecurity, as well as ethics in synthetic biology, holds paramount importance for science students. In an era characterized by remarkable advancements in biotechnology, students must grasp the significance of responsible scientific practices. Understanding biosafety and biosecurity protocols equips them with the knowledge needed to prevent accidents, mitigate risks, and protect the environment and public health. Moreover, ethical considerations in synthetic biology are essential to guide students in making morally sound decisions when manipulating living organisms. By instilling these principles early on, we empower the next generation of scientists to harness the potential of synthetic biology while ensuring that their innovations uphold ethical standards and prioritize safety, thus contributing to a brighter and more responsible future in the field of biotechnology. Holding these beliefs in mind, we create a 3-part learning packet on the topic of biosafety, biosecurity, as well as ethics in synthetic biology, suitable for students from secondary school to first years of university.