Team:St Andrews/Results
Results
HUMAN PRACTICES
Stakeholder Interviews
Through the course of our project, we made sure to meet with an abundance of stakeholders, experts, and the public. We wanted to make sure our scientific developments closely aligned with the needs of experts, latest science, and trends in the sunscreen market.
Results and Findings:
- Identified possible risks of the release of E. coli into the environment, during the meetings with Dr Nicola Allison and Prof Leslie Firbank
- Invited Prof Firbank as one of the panellists of the Synthetic Biology Forum
- The meeting with Dr Hennige and Dr Tagliati informed us on the existence of other chemicals, in addition to UV filters, which are harmful to marine life
- Idea of gene circuit optimisation – suggestion of Dr Helder Ferreira
- Discovered the possible benefit of permanent integration of bacteria in the interview with a Professor in Dermatology
- Gained insight into the industry by the interview with Cornelia Schürch and Louisa Laing
Discussion:
The stakeholder meetings had valuable impact throughout the full length of the project. The recommendations of the stakeholders were widely integrated in our modelling of the gene circuit as they encouraged us to design a killswitch, which minimises any potential risks for the environment, and to create a complex evolutionary-mutation model. Following the Dr Ferreira suggestion, we optimised the gene circuit to increase the efficiency of expression. The meeting with Dr Tagliati encouraged our team to develop the Coralware app, which unlike the existing apps is focused on environmental impact. The meetings with Professor in Dermatology, Lousa Liang and Cornelia Schürch had significant impact on the future perspective of the project.
Twitter Sentiment
The aims of the Twitter sentiment analysis were the assessment of most recent prevailing opinion towards GMO and identification of the words GMO is associated with. The analysis was conducted with a perspective of implementation in market research, education and outreach of the project, as well as, serving as an up-to-date source of information regarding the impression of GMO in the society.
Results and Findings:
- Basic Data Analysis identified top 10 most liked and most retweeted tweets – two dominant topics – global nutrition (negative sentiments toward GMO) and scientific discoveries (positive sentiment towards GMO).
- Method 1 showed the mean (0.03977) and the median (0.008657) of the score, calculated by the first method, when rounded to the integer value were equal to 0.
- Method 2 showed higher percentage of positive sentiments (24.15%) compared to the negative (15.21%).
- Method 3 produced wordclouds and tables with the most frequent words, contributing to the second aim.
Discussion:
The two aims were successfully achieved. Regarding the impression of GMO the three methods showed similar results of the prevalence of neutral sentiments towards GMO. The findings about GMO associated topics were widely integrated in the entrepreneurial aspect and human practices. Therefore, scientific significance was added as another possible focus of advertisement of the potential product. This information supplemented the recommendation of one of our stakeholders – Louisa Lang, that the emphasis of the sunscreen should not be on GMO, but on the environmentally-friendly aspect. Besides, in many tweets the adjective ‘natural’ was used as opposite to GMO and in such tweets the former was believed to be better. Therefore, we regarded this as an evidence of the meaning of the term GMO being unclear for some people and related this information to the reports from the synthetic biology forum we organised, as well as, the survey results.
Survey
The survey served to collect information about people’s perceptions of a potential new probiotic sunscreen which is genetically engineered and contains live harmless bacteria. The initial concept of the survey in terms of comparing responses from different locations was altered due to insufficient data but the final results were still very valuable. The version of the survey was different in China due to the change in the platformed used and the results were analysed separately.
Results and Findings:
- High level of environmental awareness within the participants was demonstrated
- 83.58% of the people said they use sunscreen while being at the beach; 69.57% in China
- Controversial opinion about GMO safety
- Consistency and price most important when buying sunscreen; Type and Brand in China
- Half of the participants were positive towards the use of our potentil sunscreen but expressed safety concerns
Discussion:
The results provided information on the variety of the demands of the market in different locations which could be used in the market research and confirmed the importance of killswitch incorporation in the design of the gene circuit.
Forum
At the end of August 2020 our team hosted an online Synthetic Biology Forum on the provocative topic “Will GMOs shape the future of humanity”. Five experts from different realms of science discussed this topic and lots of other questions related to their individual research, synthetic biology and iGEM.
Achievements:
- More than 2300 reached viewers on social media
- Gathered scientists from different fields
- Raised awareness towards synthetic biology and genetically modified organisms
- High level of engagement of the participants in the Zoom meeting
Results and Findings:
- 90 % of the participants said they were more interested in GMO after the forum compared to before
- 50% of the participants said they would use GMO as cosmetics or therapeutics, while the rest 50% said it depended on the case
Discussion:
The Synthetic Biology Forum had great success and the topic attracted the attention of the public to synthetic biology. The results of the feedback survey showed that the participants enjoyed the interactive event and it did influence their knowledge and opinion towards GMO. Moreover, we collected more information on the relation between informational scientific events and change in the opinion of potential customers, which was then implemented in the business potential strategies for the future of the product.
Coralware App
Achievements:
- App software development
Results and Findings:
- Current apps do not give enough details regarding environmental impact of the cosmetics ingredients
- Creation of a database of common sunscreen ingredients
- Classification of the chemicals contained in cosmetics according to their toxicity
Discussion:
The Coralware app was inspired by one of our stakeholders Dr Tagliati and it was another way of raising awareness of current research on ecotoxicity and allowing the potential customers to make their own decisions based on clear, nonbiased information.
Education
Achievements:
- Creation of lab-based video of the simplified process of soil sample analysis and the biological principles behind it
Discussion:
Developed understanding among primary school students of the complex processes involved in the soil analysis. In addition, we hope we contributed to increasing the interest of students in Biology.
COLLABORATION
Collaboration with other teams was an essential part of the human practices. Our team hosted the UK iGEM teams meet-up and collaborated with teams from China, Australia and the UK. With the Manchester and Hainan team we worked together extensively, which resulted in a mutually beneficial partnership.
Collaboration
Achievements:
- All UK iGEM teams joined the meet-up with the overall number of participants 65
- Hosted a workshop at Paris meetup due to collaboration with Bettencourt
Results and Findings:
- Recruitment of larger number of participants in the survey
- Learning more on coral conservation on the Symposium, organised by the UNSW team
Partnerships with the Manchester and Hainan teams
Achievements:
- Conduct several stakeholder interviews with Manchester
- Able to meet with a dermatologist and Mewburn Ellis patent attorneys at Manchester's invitation
- Received a certificate for participation in Market Research Seminar by the Manchester team
Results and Findings:
- Additional data about public opinion from the surveys of the Manchester team
- Safety assessment of the potential ingredients of ‘Shinescreen’ in the wetlab of Hainan team – no ingredients were found to be toxic
Discussion:
The UK iGEM teams meet-up was of a great importance for the further collaboration between the teams. We met the Manchester team and led many aspects of the human practices campaign together which made us work more efficiently and reach stakeholders that we would not be able to discover alone. The other long-term collaboration which turned out into a partnership was with the Hainan team. The wetlab experiment of Hainan served as a starting point in the creation preliminary list of environmentally friendly ingredients for our sunscreen.
ENTREPRENEURIAL
We have conducted a series of stakeholder interviews to explore our busines responsibilities, conducted market research, consulted with several IP experts to explore patenting, and successfully pitched our idea. Currently, we are at a stage where we are making a business development plan are in the process of setting up a company.
Achievements:
- Made useful connections for future business opportunities with IP lawyers, the St Andrews technology transfer centre, researchers, medical experts, and companies in the sunscreen industry.
- Won the St Andrews Enterprise week Pitch event with our business proposition. Won £980 to cover our lab costs in phase II.
- Conducted market and SWOT analyses for our unique value proposition based on direct and indirect market research
- Gained extensive knowledge on intellectual property protection.
Results and Findings:
- Found a unique market opportunity and value proposition for our sunscreen
- Found unmet consumer demands which Shinescreen could meet in different regions across the globe
- Discovered consumer hesitations about our product due to its GMO properties
Discussion:
Overall, we developed a more cohesive understanding of the Suncare market and the unique position our product could fill in this market. Our human practices greatly influenced our entrepreneurship during our project, and the market opportunities we found through our entrepreneurial work can influence the direction we pursue with our phase II wetlab work.
MODELLING
Gene circuit and Biosafety
To pursue the prospect of ‘Shinescreen’, The St Andrews iGEM Team of 2020 has harnessed a failproof dual plasmid circuit design, consisting of two components. The first component is called ‘shinogen’ and is responsible for the synthesis of shinorine. In terms of biosafety in order to study the evolutionary stability of the killswitch, we devised an evolutionary mutation algorithm we called the ‘Genetic Algorithm’ to test the resistance of the second component thanogen specifically to point mutations across n generations.
Achievements:
- Failproof dual plasmid gene circuit deign
Figure 1. Finalised Gene Circuit
- Maximising ‘shinogen’ efficiency by knockout of pentose phosphate pathway genes and codon optimisation of pre-existing non-native parts by IDT codon optimisation tool
- Creating killswitch mechanism – ‘thanogen’, Incorporation of UV sensing system and glucose regulatory activation
- Optimisation of BioBrick RFC [10] and [1000] by in-silico site-directed mutagenesis
- Added information on existing parts in the Standard Biological Parts Registry based on literature review
Results and Findings:
- The ccdAB operator/promoter (BBa_K3634013) was predicted to have 5 binding sites for ccdA2 controlling expression of Lon protease, later replaced by mf-Lon as Lon protease is already native to E. coli
- Predicted that an RBS existed after the ccdAB promoter region
- Fully neutralise leaky protein expression of mf-Lon protease by additional weak RBS
- All constructs of the plasmid after digestion with the selected endonucleases were found in silico to have been suitably fragmented to prevent HGT of active parts:
Figure 2. Molecular weights of pSB3E1-AN (lanes 1,2,3,4), undigested (lane1), digested with R.CviJI (lane2), RFC[10] illegal endonucleases (lane3) and RFC[1000] illegal endonucleases (lane4). Molecular weights of pSB3B1-DOPH (lanes 6,7,8,9), undigested
- Showed that thanogen which is more resistant to evolutionary drift with transient point mutations in one of the single toxin/antitoxin non-functionalising the killswitch than the just a ccdA/ccdB system alone.
- The present thanogen showed greatest sensitivity in the ccdA-ccdB binding and protease activity, likely on account of the higher likelihood of mutations occurring within the CDS, which comprise the longest number of nucleotides
- The design of the killswitch ensures that the default state of the cell is cell death: dysfunction in protease, ccdB and endonuclease must arise before cell stasis/death occur but most drift resulted in a compromise to cell life.
Discussion:
Through human practices activities, it became clear that the biosafety of our product was a key concern. Escape into the marine environment or permanent integration into the skin microbiome are the two considerable possibilities that we wish to avoid, both in terms of escape of the bacteria themselves or escape of the genes in our circuit by horizontal transfer. We decided on a two-plasmid design, splitting the shinorine producing genes between both plasmids. We also split the components of the kill switch between the plasmids in such a way that loss of one plasmid results in cell death. This minimises the risk of our genes being horizontally transferred into the native microflora, as uptake of either plasmid without the other will be detrimental.
By updating the description of pre-existing parts we would facilitate future users to have better understanding of the system. We have also contributed to the second phase of the project in 2021, where the dual plasmid design would be implemented.
The modular nature of our gene circuit will allow future transfer of components such as the kill switch and elements of the shinogen to different probiotic cosmetics and therapeutic products. In-vitro testing in 2021 will assess and inform the modelling assumptions made this year and allow for promoter and RBS optimisation to maximise the efficiency of our genetic system.
Macroscopic Modelling
In order to investigate the behaviour of our chassis organism once applied on the skin we worked on macromodelling.
Achievements:
- Creation of ‘Macroscopic Skin Model’ – spatio-temporal model which determines how several relevant macroscopic variables evolve on the skin after application