Team:Linkoping Sweden/Human Practices
Human Practices
Human Practices is an essential part of any iGEM project. From the official site of iGEM:
We have put considerable effort and consideration on the environmental implications, social effects and future real world implementation of our project. This has resulted in a desalination system that we believe has the potential to improve the lives of many people worldwide. Read more about our approach to sustainability here.
Integrated Human Practice
iGEM teams' Integrated Human Practices efforts take many shapes and forms, but the goal is always to interact with experts in the field and people that are somehow affected by a problem related to the project. Since our iGEM team has designed a desalination method that uses modified phototrophs, our focus has been to reach out to people related to freshwater scarcity, cyanobacteria and the specific proteins we want to introduce in our modified organisms.
Water scarcity
Lack of freshwater, known as water scarcity, is one the biggest threats to global health. Even though 70% of the planet's surface is covered in water, only 2.5% of that water is freshwater. And out of the total freshwater, only 1% of it is accessible for human use. This leaves approximately 7.9 billion people with only 0.025% of the total water resources [1]. Out of the freshwater, 70% of it is used for agricultural purposes [2]. The Global Water Institute estimates that 700 million people could be displaced by severe water scarcity by 2030. This is further complicated by globally increasing temperatures due to climate change [3].
CyaSalt, which is our environmentally sustainable solution to desalinate water, could theoretically have a largely positive impact on this global issue if upscaled to a functioning industrial level. Therefore, we wanted to learn about the subject from people in the academic world, and interact with people in the industry.
Dr Sara Ahmed is an experienced Adjunct Professor based at the Centre for Water Research, Indian Institute for Science Education and Research (IISER) Pune, India. She has over 30 years of experience on the socio-political economy of water in India and is the founder of the Living Water Museum, a virtual repository visualising narratives on water heritage (built, natural, cultural). Sara has also been a part of multiple international water-related projects and is on the board of WaterAid, India and Wetlands International South Asia, as well as the Global Network of Water Museums. In a collaboration with the iGEM team from IISER Pune, we got recommended to contact her since our desalination project is heavily connected to water and the global issue regarding scarcity of it. We learned from Sara that in India, a lot of investments go into the water cleaning industry, due to the hardness of the water in semi-arid regions of the country, such as Gujarat where Sara worked since 1992. To counteract this, a reverse osmosis system is utilized commonly by many households. An issue with this method is that it filtrates different types of salt, which includes the ones that can be beneficial for the human body. According to Sara, desalination plants today can create toxic waste products. Our project idea of using modified bacteria instead provides a more eco-friendly solution, and could be a good solution according to Sara, if it’s proven to be a functional system. Desalination plants also use huge amounts of energy, which costs a lot of money and thereby affects the people in parts of society with a low socioeconomic status the most. Our project proposes a solution that requires far less amount of energy to desalinate the water. Shouldn’t everyone have the right to clean water?
Farmer's perspective
To get a better perspective on how water scarcity affects people here in Sweden, and to get advice on how we should design our desalination system, we visited farmers on the island of Gotland situated in the Baltic Sea. You can read more about the design we made for our future implementation on our Implementationpage. Gotland is one of the few places in Sweden where water scarcity is a real, very present, problem. We reached out to and met four different farmers on their farms for in depth discussions. The farmers we met were Rune Hägg, Håkan Karlsson, Karolina Pettersson, and Göran Berntsson. They do slightly different agricultural work but have all been experiencing hardships due to lack of freshwater. When discussing how water scarcity has affected them, and what they do to counteract it, they told us that due to droughts on the island, many farmers on Gotland have had to ration their water and let a lot of crops dry out and die during dry periods. The water scarcity also affects the cattle, leading to a shortage of grown cow fodder as well as a lowered milk production from the cows, something that largely affects the farmer’s economies. We also learned that cows are even more dependent on clean water than humans because they run a much higher risk of dying when drinking water that is contaminated with, for example, E. coli. To adapt to the droughts, some farmers have built special dams, called environmental ponds, that collect water throughout the year. This also enables the farmers to catch fertilizer before it reaches the sea. These dams can provide up to 12 farms with water, but sadly, not everyone has access to them. Furthermore, they have dried up during particularly warm and dry periods.
With the summer temperatures increasing year by year, water scarcity is only getting more severe. Watering bans have become more common, and according to Rune Hägg, many farmers are worried about their water supply running out. In 2018, Sweden experienced an exceptionally warm summer, with some places reaching up to 33 ℃. This led to some farmers, who usually grow their own cattle fodder, having to order fodder from the mainland, costing them an extra 60 000-70 000 €. The economic effects of this heat can still be felt to this day for many of the farmers.
Overall, the farmers had a positive attitude regarding our project. However, one of the farmers expressed slight concern regarding the usage of GMOs saying:
Another farmer was very positive toward our project and said that he firmly believes that GMOs are the future for agriculture and that it needs to be accepted and understood by the public.
The consensus towards the use of GMOs was that, as long as it has been tested and approved by governmental and other institutions, they would consider using our product. However, the most important factor that all farmers had as their main concern was the cost and the fact that they preferred a local setup. The farmers were very clear that they would only purchase our product if it was a more affordable alternative to either getting their water from another source or reducing their agricultural output. Therefore, our team started putting more effort and thought into how to make our theoretical industrial setup as affordable as possible, while at the same time upholding a good desalination standard that could provide farmers with usable freshwater. It also led us to focus more on designing our up-scaled desalination set-up in a way that is well-suited for farms situated by the coast. For a more in depth explanation of this theoretical industrial set-up design, visit our Implementationpage.
The cyanobacteria - Synechosystis sp. PCC 6803
Cyanobacteria is a so-called phylum (the classification between kingdom and class), of photosynthetic bacteria. They live in aquatic or moist environments and are believed to be the first organisms on earth to develop photosynthetic properties [4]. Like other phototrophic organisms, cyanobacteria utilize sunlight to fixate carbon and convert it into oxygen, among other things. One commonly used cyanobacteria for genetic engineering purposes is Synechocystis sp. PCC 6803. It was also the first phototrophic organism to have its genome completely sequenced [5]. Since none of the people on our team had previous experience working with Synechocystis sp. PCC 6803, or phototrophic organisms in general, we were in need of experimental advice early on in our project. Therefore, we reached out to the scientific community in different ways.
The Experts
Dr Vamsi Moparthi is a principal research engineer at Linköping University. He is currently researching Myc proteins, universal upregulators of gene expression, and their role in cancer development. Before joining the research group in Linköping, he worked with cyanobacterial genetics at both USC Berkeley and Uppsala University. You can learn more about Vamsi’s current and previous research by listeningto the 5th episode of our podcast, Attempting Science. Due to Vamsi’s extensive work with both Synechocystis sp. PCC 6803 and Synechococcus elongatus, as well as the fact that he works in a lab next to ours, we decided to contact him to get some help on cultivating and transforming Synechocystis sp. PCC 6803. We received protocols for both cultivation and transformation, as well as practical tips regarding the light intensity and cycle. Vamsi also provided us with some articles related to vectors we could use for transforming our target genes into Synechocystis sp. PCC 6803.
Dr Julie Zedler is a junior professor in Synthetic Biology based in Jena, Germany, with a special focus on photosynthetic organisms like cyanobacteria. Julie has been active in different seminars and discussions forums regarding phototrophs, and this is where we first came into contact with her. She thought our project sounded ambitious and believed that we would later encounter problems in experimental parts, which came to happen. Our plan of cloning multiple proteins into the bacteria Synechocystis sp. PCC 6803 could, according to Julie, possibly change the metabolism of the bacteria and have a high risk of failing due to complications with protein folding. This could support the idea of using some other phototroph species that could sustain synthesis of such complex proteins. Since Julie has been working with similar strains for multiple years we also got practical lab advice, like how to avoid plate contamination, a proven transformation protocol for transformation of Synechocystis sp. PCC 6803 and how to modify the growth media for optimized bacterial growth.
Dr Paul Hudson is an Associate professor at KTH, Stockholm and the Principal Investigator at the Hudson Lab, Science for Life Laboratory. His research concerns metabolic engineering of carbon fixating phototrophic bacteria, Synechocystis sp. PCC 6803 for example. We reached out to Paul due to his experience with Synechocystis sp. PCC 6803 and were fortunate enough to get a sample of wild type Synechocystis sp. PCC 6803 to use for further cultivation in our wet lab part of the project. After we pitched our project idea, Paul was positive about our chances of successfully expressing halorhodopsin and CBD in Synechocystis sp. PCC 6803 and also agreed upon that the strain probably was the best choice for iGEM purposes. Another option would have been Synechococcus sp. PCC 7002, but we decided not to use that strain. We also discussed plasmids and he agreed that our choice of using pEERM1 and pEERM4 was good for the purpose of integrating the cloned fragments into the genome of the host bacteria. We also got all kinds of practical advice for the wet lab, like incubation conditions and what kind of carbon energy source to use for the carbon fixation of the cyanobacteria.
Magnus Refthammar is Director of Product Development at Dynamic Code, a growing HealthTech company based in Linköping, Sweden. Magnus is an alumnus from Linköping University with a degree in Engineering Biology and is also one of the founding members of LiU iGEM. The team participated in iGEM for the first time in 2013. Since Magnus is passionate about LiU iGEM and the possibilities of synthetic biology, he was eager to contribute to this year’s project, CyaSalt, in some way. Magnus visited the 3rd episode of our podcast, “Attempting Science”, and talked about his iGEM experiences. We also discussed industrial upscaling and Magnus, with his experience from the biotech industry, insisted on the importance of being ambitious and thinking about long-term prospects when doing an iGEM project. The application and the industrial possibilities the project could lead to should never be underestimated. Magnus was also interested to hear about how we wanted to commercialize our project idea. We learned a lot, both on the podcast and during talks surrounding the actual recording.
Dr David Drew is a Professor in Biochemistry at Stockholm University, which we contacted regarding questions about membrane proteins and ion channels, since our project is heavily dependent on functional proteins in the membrane of Synechocystis sp. PCC 6803. We learned from him that it’s not possible to accumulate an ion gradient with only non-selective ion channels activated by osmotic pressure, since this will result in a strive towards thermodynamic equilibrium. This made us consider our parts and led us to the path of using channelrhodopsin, since it functions in a similar fashion to halorhodopsin but can import sodium ions, which we believe will balance out the negative gradient accumulated by the import of chloride ions by halorhodopsin. From this, we also decided to use MscL to hinder osmotic pressure (from the ion influx) to cause cell disruption.
[1] National Geographic. Freshwater Crisis. Cited 2021-10-16. Available from: https://www.nationalgeographic.com/environment/article/freshwater-crisis
[2] OECD. Water and agriculture. Cited 2021-10-16. Available from: https://www.oecd.org/agriculture/topics/water-and-agriculture/
[3] UN water. Water scarcity. Cited 2021-10-16. Available from: https://www.unwater.org/water-facts/scarcity/
[4] TBiologyOnline. Cyanobacteria. Cited 2021-10-16. Available from: https://www.biologyonline.com/dictionary/cyanobacteria
[5] Amezaga JM, Amtmann A, Biggs CA, Bond T, Gandy CJ, Honsbein A, Karunakaran E, Lawton L, Madsen MA, Minas K, Templeton MR. Biodesalination: a case study for applications of photosynthetic bacteria in water treatment. Plant Physiol. 2014 Apr;164(4):1661-76. doi: 10.1104/pp.113.233973.
[6] Ikeuchi, Masahiko & Tabata, Satoshi. Synechocystis sp. PCC 6803 — a useful tool in the study of the genetics of cyanobacteria. Photosynthesis Research
volume 70, pages 73–83 (2001). Cited 2021-10-16.