Human-centric Integrated Approach

Our team has believed from the very beginning that iGEM projects are for the greater good of humanity and this planet. Every iGEM project is inspired by real problems and people; so is ours. The aim of every iGEM project is also to inspire many more to work for the greater good of society through science, research, communication and collaboration. We wanted our project to target a global problem and try to solve it using a local solution along with the magic of synthetic biology and we believe this encompasses the beauty of our project.

Our project - ‘JetroEco’ is addressing the effects of climate change and how we can try to mitigate them using our knowledge and engineering in synthetic biology. Our team had multiple ideas that were extensively discussed during the months of February to April, before finally arriving at JetroEco.

• February

  • Drug delivery systems and ADCs (Antibody Drug Conjugate) - We were looking into how we could harness the immense specificity of antibodies to try to treat diseases like Cancer where specificity is of utmost importance.
  • Aptamer-based therapeutics - Foot and Mouth disease is a major killer in the cattle industry, which causes a huge economic loss every year. Our approach was first to diagnose it with the help of aptamers and simultaneously treat it.
  • CO₂ reduction - The world is at a stage where we need to take strict measures to reduce CO₂ emissions. We were thinking of various ways to do this. One of the apparent ones was carbon capture in modified organisms to produce industrially useful chemicals.
  • Cancinerf - We tried to look into chemotherapy's side effects and mitigate that via the NRF2 pathway.
  • Synthetic cloth degradation - Fast fashion, which is an unsustainable way of producing clothes, has unfortunately seen an enormous boom. We were planning to engineer organisms that could degrade these materials efficiently.
  • Bioremediation - One of the members was very enthusiastic about working on using Chlamydomonas reinhardtii and engineering it in a way such that it can capture heavy metals and fix them in an alginate bead. However, there were issues with making a biofilm for the same.

• March

  • Drug delivery systems and ADCs (Antibody Drug Conjugate) - The idea progressed to a stage where we started thinking of making a system where the drug (a natural compound like Paclitaxel), along with the linker and Antibody, will self-assemble to form the ADC construct. However we dropped the idea owing to its complexity and feasibility in the iGEM time frame.
  • Aptamer-based therapeutics - The idea eventually died out when we spoke to a few experts and learned that the government has already invested lots of money into compulsory immunisation against FMD at a very cheap cost with which we won’t be able to compete.
  • Biofuels - From the CO₂ reduction motivation, we were looking into various industries with a heavy carbon footprint, and we came across the transportation sector, which accounts for about 1/5th of all global greenhouse gas emissions.
  • Biomanufacturing of natural products of importance - It came as an interesting field where we were looking for various materials which are of use and could have problems with natural production. A few materials we were looking into were terpenes, rubber, cumin etc.
  • Cancinerf - The idea went forward and saw more developments with regards to the dry lab where the team members had figured out which genes were to be targeted and started to design a peptide to do so.
  • Sandalwood Spike disease - India is home to a huge number of top-quality Sandalwood trees and this was threatened by a phytoplasma disease called Sandalwood Spike disease (SSD). We started looking into the literature and spoke to a few experts.
  • Asthma- One of our teammates was looking into asthma and its treatments. They thought to block IL8 which is responsible for the inflammation using GEMs but because of its possible off-target effects, the idea was discontinued.

• April

  • JetroEco - We finally came up with a fully furnished idea after we came across this interesting article on a test flight using Jatropha oil back in 2018. As we were already working on Biofuels and trying to produce natural substances along with the aim of CO₂ reduction, this idea perfectly fit. By the end of the month, this came up being the selected one we wanted to proceed with. Lots of developments were being made recently in the area of SAFs so we thought it will be a nice opportunity for us.
  • Cancinerf - The idea was very well planned out, however, we felt that it was a little tricky because of the magnitude of the problem we were trying to solve. There were other logistical reasons as well to drop the idea.
  • SSD - This idea also came to an end as there was very little literature about SSD. We got to know that it takes a lot of time to work with woody plants.

JetroEco was inspired by an old news article from 2018 which covered India’s first ‘Green flight’ test run from Dehradun to Delhi by the airline SpiceJet. The aviation sector has been a major stakeholder in global greenhouse gas emissions, with around 1 billion tonnes annually, and this figure is set to increase with increasing globalisation and and utility of air transport. We have come a long way since the first flight flown by the Wright brothers in 1903, both in terms of technological advancement and environmental impact.

When the world is seeing a shift towards electrification of the transport sector, aviation and marine transport seem a distant dream ^[1]. To reduce the carbon footprint of the aviation sector while keeping in mind its potential growth, biofuels or Sustainable Aviation Fuels (SAF) seem to be the best bet for now. This is important for our country India because it is projected to become one of the largest shareholders in the aviation sector ^[2]. To learn more about the project visit our project description page.

While designing the course of our Integrated Human practices, we kept the above design principle in mind. We have tried to make our project as broad as possible and also reach the depth of every activity we have undertaken. Our approach has always been very human-centric while also keeping in mind ethical and moral considerations. All the human practices activities have been undertaken after seeking consent from the respective individual.

Understanding the Problem

One’s iGEM project can be understood with the help of people associated with the problem, taking their input on the existing situation and advice on how to make it better. For JetroEco we have done a Stakeholder analysis to identify different sets of people who will be affected and associated by our actions. JetroEco has tried to reach out to a diverse set of stakeholders and reflect on their suggestions.

There is no doubt about the effects of climate crisis. After our project to work on Sustainable Aviation Fuel (SAF), we started looking for various candidates for our SAF. The first ‘Green’ SpiceJet flight used 75% Aviation Turbine Fuel (ATF) along with 25% oil from Jatropha curcas seeds (after Hydroprocessing) in one of their engines and this was shown to reduce CO₂ emissions. We then researched on Jatropha curcas plant and its oil and found out that it is one of the best to be used as it doesn’t require much processing (used as Drop-in fuel) and it complies with all the standards of American Society for Testing and Materials (ASTM).

Then we started looking for Jatropha curcas cultivations and how its oil is produced. To our surprise, we learnt that there is barely any Jatropha cultivation in India which is supposedly the largest producer of the oil. This means that there is negligible supply of Jatropha curcas seed to produce biodiesel despite being the best candidate. We tried to understand the cause behind it from our different stakeholders.

Ideating Our Solution

Values and Framework

After carefully analysing the inputs and expectations of all our stakeholders, we decided to try to add various aspects to our project design. We set out to produce a SAF that is environment-friendly and economical. Along these lines, our SAF will be:

To achieve the above goals, we developed the project in a scientifically robust way but taking expert advice from multiple people all across the world.

Synthetic Biology

The need for SAF and the failure of a suitable feedstock for the same led to the inception of JetroEco. We felt that Jatropha curcas oil, despite being the best candidate for SAF had a lot of limitations in terms of growing the plant, so why not harness the magic of Synthetic biology to mass-produce Jet fuel that is derived from Jatropha curcas fatty acids?

This is when we started looking into various research papers and found that Jatropha curcas' Thioesterases - JcFATA and JcFATB are responsible for the fatty acids in the plant. We found one paper that tried to express these thioesterases in E. coli to change its fatty acid content to the one of Jatropha curcas ^[4]. We started looking more into this idea and eventually decided to try to express Jatropha curcas thioesterases - JcFATA and JcFATB in E.coli and change its fatty acid pool to produce Jatropha curcas triacylglycerol (TAGs). This was our starting point and it saw immense developments with the inputs of various experts in academia and industry.

Implementation and Evaluation

After careful consideration from all our stakeholders and experts, we were finally able to design our project and execute our wet lab and dry lab experiments and collect data. It is important for anyone in the scientific community to carefully analyse the data using various qualitative and quantitative tools.

Proof of Concept

To understand if our thioesterases have been expressed and are producing our desired fatty acids and hydrocarbons, we need to quantitatively measure that using sensitive techniques like LC-MS and GC-MS.

We ran our samples in the LC-HRMS and the GC-MS facility at the National Chemical Laboratory (NCL) in Pune. For LC-HRMS for quantification of our fatty acids, we were assisted by Dr Neelakshi Sadavarte and her PhD students - Megha and Mridula, and Aakash C - our PhD mentor in analyzing the data obtained. Dr Sanjay Borikar helped us with GC-MS for detecting the hydrocarbons in our samples.

To find all details, visit the Proof of Concept section.

Bioreactor Design and Entrepreneurship

Once we have our engineered strain producing enough amount of our desired product, we need to scale it up. For this purpose, we need a bioreactor of higher volumes as compared to conical flasks in the lab.

Dr. Anand Ghosalkar

He suggested the various parameters we need to keep in mind to design a bioreactor for our project. He also explained us the process of glucose extraction by pre-treatment, something that even we could do to obtain our media at a much lower cost.

He is the Chief Scientist at Praj Industries and an expert in biochemical engineering, research on renewable chemicals and materials and second-generation biofuels. To understand the details of the bioreactor, visit our hardware page linked here.

Dr. Siddhesh Kamat

He is an associate professor at IISER Pune working on Chemical biology, metabolomics, chemproteomics, lipid metabolism, and protein biochemistry.

He also said that cost is an important factor that should be taken into consideration during the manufacturing process. As sustainable fuels are at a much higher cost, research should try to reduce it in order to make it economically viable. This gave us the idea of trying to analyse the cost of our product using our kinetic model and its output. For the cost analysis, visit our entrepreneurship page linked here.

Policy Making and Governance

Many times we have the technology and skills to bring about a major shift in society, however, the impetus is received only when there are appropriate policies in place to ensure that the purpose is met. Even though the general public, airline companies and fuel companies are willing to shift towards SAF, but there are currently no policies to ensure mass production of the same. This is a hindrance to the development of SAF and unless we have a push from the government, it is difficult. If there is soon going to be a mandate of 1% SAF blending, then we need more incentives to produce SAF and reduce its cost with the help of research.

Prof. Carlos Henrique de Brito

Dr. Carlos is a member of the Brazilian Academy of Sciences, Senior Vice President (Elsevier), and Professor Emeritus; Elsevier Research Networks; Physics Institute, University of Campinas. We had met him during our poster presentation at the G20 summit held at our institute IISER Pune.

He was particularly impressed by our project so much so that he praised us on the G20 stage in front of delegates all over the world which was truly an spirit-lifting moment for us. Here is the video:

We got the chance to interact with him more and learnt about the Bioeconomy of Brazil which stands as a true example of the excellence of government policies that helped in achieving energy sufficiency in Brazil after World War II. Due to government incentives in Brazil, the farmers started mass-producing sugarcane, and a lot of this was used in the production of bioethanol. The objective of the project was to achieve self-sufficiency in energy but it came with the additional benefit of having a lesser carbon footprint.

He had worked on the topic of biofuel himself and he believes that it is the key to breaking the monopoly of the crude oil market.

His suggestions inspired us to talk to more environmentalists and policymakers to start a dialogue on carbon emissions and sustainable energy sources. This was done by bringing together a diverse set of researchers and policymakers on a common platform and taking their opinions. We organised a panel discussion and got inputs like:

• Achieving the long-term goal of net zero emissions can be started with lower emissions and balancing minimal positive emissions with possible carbon sequestrations.
• Even though the production of biofuels from biofuel crops is less carbon emissive, this could lead to a conflict of fuel over food. The panel agreed that the evolution of science and technology can reduce net carbon emissions.

Our experience at the G20 summit was enriching where we presented our project and got a chance to interact with many delegates from different countries like Mr. Casper Mayer (Germany), Mr. Piers Purday (United Kingdom), Dr. Carlos (Brazil), scientists and businessmen like Dr. Ravindra Utgikar (Praj) and even ministers like Dr. K Sanjay Murthy.

Closing the Loop

After identifying our stakeholders, we had enlightening interactions with a diverse set of people who are directly and indirectly related to our project; and whose inputs helped us shape the course of our project. The suggestions from various experts in both experimental and computation design have been a blessing, without them we would not have been able to achieve what we could.

Now it was our turn to return our share to the society. This is not only by trying to manufacture a sustainable bio-jet fuel with much lesser carbon emissions, and good performance at a reduced cost but also to communicate and educate them about the beauty of synthetic biology. We have made an honest attempt to try to reach out to all sections of the society through carefully designed activities which you can find on our Education and Communication page which is linked here.