FIAT LUX is solving one of the main problems in bacterial
observation: how to track bacteria efficiently AND over time in tissues without collecting
samples. Indeed, methods available today to track bacteria don’t have both features. In plants:
The fp genes can be used to observe bacteria throughout time but the main problem is that organisms have autofluorescence which interferes with the FP proteins, reducing the efficiency of the observation.
Another existing method is the Patho test which consists in cutting and crushing the leaves. This is very efficient but can’t be used over time.
Visual analysis can be used to monitor an infection throughout time but with low efficiency because the symptoms do not always correlate with the pathogen’s position.
FIAT LUX overcomes all of these issues as it directly targets the
pathogen, it lasts over time, and it is effective in situ since living organisms have
autofluorescence but no autoluminescence (Figure 1). In that way, phytopathogens can be tracked and the
evolution of the infection can be monitored.
Figure 1 - Comparison of existing solutions with FIAT LUX for the observation of pathogens throughout time
Analyzing what the interests and powers (e.g. means of action) of the stakeholders of our product are, is key for the construction of our business plan in order to meet their needs and requests. We divided our stakeholders into three categories: farmers (yellow dots), research centers (blue dots) which are either in agro-industries or in academies, and public institutions (purple dot) in the Figure 2.
Farmers have a big interest in our product. Indeed, they are the
first to directly face crop losses due to phytopathogens' propagation. Our tool will allow
researchers to find new treatments for epidemics which means less crop losses for farmers, hence a
reduced economic loss for them. However, their power is relatively weak because they do not use our
tool directly and mainly depend on research innovations. Nevertheless, farmer federations
also have a big interest in our product and they are more likely to use it directly or through
research laboratories, while having a bigger financial power than individual farmers.
The private research centers have the greatest power because
of their R&D budget. Nevertheless, agro-industries have a stronger budget than the academics
research field as the final objective isn’t the same. For academics, research will allow
them to understand the infection pathway, document it and try to find a way to reduce or stop
disease. In the private sector, FIAT LUX would allow them to design new products against
pathogens, tools or methods in order to generate revenues.
Finally, public institutions such as States and unions of
States (e.g. European Union, WHO) have mid-range interest for our product because it may lead to
the drop of crop losses which ultimately can help solve hunger, which is the second sustainable
goal of WHO for 2030. However, in order to succeed at this task, FIAT LUX may be one of the
solutions with a lot of other technologies and measures they may be interested in. Their power
is also mid-range because they will not use our product directly but can give fundings and allow
grants to other stakeholders to use FIAT LUX or to our company to accelerate research in
order to offer an improved product.
Figure 2 - Stakeholders analysis
We analyze our business model and core competency in the case where
we sell E.coli MFDpir transformed with our plasmid pSEVA521-fiatlux or
pSEVA531-fiatlux containing our biobrick, in the SWOT matrix below (Figure 3).
Figure 3 - SWOT matrix, in the case where the business model is to only
sell the biobrick
Thanks to the SWOT matrix, we can identify two big weaknesses:
the fact that the client needs to invest in R&D to use our tool, and that our tool
necessitates the use of an expensive hardware composed of a CCD camera (we used a ChemiDoc XRS+, Bio-Rad which costs ~$15,000 (Bio-rad 2022)). The first weakness can be solved if we handle
the R&D part of choosing the right plasmid and doing the cloning and
transformation/conjugation for the clients. The second weakness can be solved if we build an
appropriate hardware device ourselves, that is cheaper than the ones currently available on the
market. We understood that this was an unmet need in the field of synthetic biology: a simple,
cheap and feasible device to analyze bioluminescence with a user-friendly interface. The
luminescence produced thanks to our tool is important enough to be detected with a simple
smartphone. With the support of Kubii (an electronics supplier we approached), we were able to design and
construct our hardware connected with a software we already developed.
(*): in the contract form we will specifically explain that performing a
conjugation between two different pathogenic strains is prohibited.
Figure 4 - SWOT matrix, if we adjust our business model (by selling
pathogens transformed with the biobrick and providing cheaper hardware). Legend: The purple
and orange X symbolize respectively the loss of threats and weaknesses, while the blue and
orange √ symbolizes respectively the appearance of strengths and weaknesses.
Thanks to the SWOT matrix, we can see that commercializing the
pathogen directly transformed as well as providing a cheaper hardware will give us a better
competitive advantage. On the one hand, we can target more clients because there will be
no R&D needed for the adaptation of our biobrick to the pathogen wanted and a smaller expense
for the highly sensitive hardware, which can be beneficial for small structures. On the other
hand, we will be able to take a leader place on the pathogen market in constructing a portfolio
of bioluminescent pathogens. Selling or renting our hardware is also a new source of income for
our company.
The feasibility of building cheaper hardware was validated by our
team. Indeed, we successfully built a hardware for less than $400 of materials (See Hardware for details). With the
time needed to assemble the hardware, and in order to scale the production, the final price
would be doubled. In this way, we will be able to sell an efficient hardware for $800, which is
nineteen times cheaper than the device we used. The software developed will be sold with the
hardware for a total of $1,000 in order to cover the work on it and future updates.
Thanks to the market analysis, we will define our business as “commercializing bioluminescent phytopathogens for research purposes” with the possibility of buying our hardware and software.
Food safety is becoming a more and more important challenge. Companies, governments and international institutions are looking into the subject and releasing more and more funding in order to limit crop losses. During the fundraising process, we came in touch with several companies and associations in order to cover the expenses of our project. We pitched our project to them, and it triggered strong interest and enthusiasm. Two of these companies, inov3PT (FN3PT research entity, the French agricultural professional organization for potato seeds) and bioMérieux (a diagnostic company), gave us important financial support that allowed us to conduct our experiments throughout the entire project. The support of inov3PT gave credibility to FIAT LUX. We also received the technical support of many other biotech companies (Promega, New England BioLabs) that sent us valuable laboratory equipment. Companies outside the field of biotech also supported us. It is the case of Kubii, an electronics supplier, that provides us with electronic devices for our hardware. They were highly interested in our tool, as this showed how their equipment could be used in the synthetic biology industry. We also approached associations (ABIL) and foundations of our schools (FSDIE Lyon 1, INSA Lyon) that showed great interest in our project and supported us. At the same time, we organized a crowdfunding campaign that was successful as we received more than €5,000 in only 30 days. It underlines the strong interest for our solution from the public as well.
We drew up a LEAN business plan (Figure 5) in order to formalize our project in
a simple manner. It allowed us to visualize our business main components and identify the potential of FIAT LUX
and weaknesses we could then address. It is not a frozen canva but an ever-moving tool that we are
perpetually adjusting and refining.
Figure 5 - LEAN business plan
In the first instance, our team has full ownership of the
intellectual property thanks to the filling of a “national laboratory notebook” (“Réseau CURIE”
2022). This allows us to fully dispose of our tool, our research work and our innovations.
However, since our project is of great interest to one of our sponsors (inov3PT), they have
offered to incubate the project and work closely with our team. For these reasons, inov3PT’s
intellectual property over our project will have to be discussed after the iGEM competition.
We analyzed the risks our project may face using the FMECA
(Failure Modes, Effects and Criticality Analysis) method. We evaluated these risks based on
their frequency (F), criticality (C) and detection index (D). Then, we calculated the risk
priority number (RPN) in order to know the relative importance of each risk.
Regulatory risks: Because we are creating and selling GMOs, our business is subject to actual and future legislation in France and the European Union. Fast swift legislation is an actual threat and must be followed carefully.
Infrastructure risks: Our activity needs to have particular infrastructure such as BSL-2 and even BSL-3 labs and greenhouse, depending on which pathogens we are working on. Dysfunctions in the infrastructure (ventilation failure, etc.) will represent unexpected expenses and our work will be stopped until its repair, which may generate problematic delays.
Market risks: The emergence of new competitors is a threat for our business. However, this emergence is limited by the fact that our market is a high-barrier technology market (infrastructure needs).
Environmental risks 1: The production of bioluminescent pathogens will generate biohazard wastes that will need to be treated rigorously.
Environmental risks 2: The leak of pathogens outside of our securised labs, greenhouse, or during the sending to our clients will have a huge impact on the environment and public health.
Project management risks: Even if we were helped by our PIs and will be by the FEE (Student Entrepreneur Program of INSA Lyon) (see “Timeline and growth”), our team is composed of young and low experienced entrepreneurs. It may result in increased risks linked to project management aspects: lack of communication, lack of planning, under-optimized use of human resources and mistakes young entrepreneurs are more likely to do. Learning quickly from our mistakes will be key in mitigating this risk.
Table 1 - Risk assessment of FIAT LUX using the FMECA method
Thanks to the FMECA method (Table 1), we know that the biggest risk for our
business is the biohazard treatment, followed by the project management. Knowing that, we will
be very vigilant facing these two risks.
With the expansion of our business, we will send GMOs (bioluminescent phytopathogens) to our clients. Special care for safe sendings is needed. Our products will be considered as “UN 3373 biological substance category B” because they are not life-threatening to humans and animals but are pathogens. The packaging for its sending will include leak-proof primary and secondary receptacles, an absorbent material between those two receptacles, and a rigid outer packaging. The product will be sent at -80°C to prevent any alteration of it. Thus, dry ice will be placed between the secondary and the outer packaging. For the safety of the sending, the symbol presented Figure 6 shall be placed on the packaging with the description “UN 3373 Dry Ice”.
The commercialisation of our hardware doesn’t generate any safety
measures.
Figure 6 - Symbol shown on the package, diamond-shaped mark for UN 3373.
(From UCL 2020)
We actually plan on commercializing our tool. We already identified
our first customers, inov3PT who accepted a close collaboration for the
continuation of the project. The first step will be to take our research from a student project
to an actual company that we would name FIAT LUX. From there, in parallel to our lab work
(both wet and dry) and research we plan on applying to the FEE Up program incubator (“LA
FEE - INSA” 2022), which is a program run by the INSA Lyon. The program lasts 10 months and
provides business support to engineers in order to make them understand and assume the risks
inherent in the company. From the creation of our start-up to the commercialization of our tool,
we foresee the following evolution:
Figure 7 - Timeline of our project after the iGEM competition
Note: Through the incubation of our start-up in the FEE program, we will
create its legal status. We will choose to create a SAS (“Société par Actions Simplifiées” -
Simplified joint-stock company) because of its juridical flexibility and its ease of creation
(Facon 2016). It also requires little funding (minimum of $300) which suits our current
situation.
Our fully developed solution has a long term positive impact as it permits to track
pathogens throughout time and helps the design of treatments preventing crop losses.
Nevertheless, even if we work really hard to make our design solution accessible to the greatest
number of labs, our hardware, software and tool still represent a certain cost which limits its
democratization. Furthermore, our tool is a hope for farmers to see their crop losses reduced or
even eliminated, but they still need an intermediary, labs for example, to have access to the
solution provided by our tool and thus are dependent on the work of agroindustries.
In the very long term, we are considering extending our field of action to the study of pathogens in various controlled environments such as soil and water (in isolated and secured field batches to ensure that our bacteria can not interact with the exterior environment) in order to understand the mechanisms through which pathogens reach plants in such environments; that would allow labs to act and treat plants earlier in the process of infection. Another axis would be the variety selection research. In fact, some varietal phenotypes are more likely to develop some infections than others: our tool could also help to identify the phenotypes predisposed to certain diseases. More details about these aspects are presented in the Human Practices page.
Our proof of concept was done on Dickeya solani.
This phytopathogen is responsible for important crop losses all over Europe on potato fields.
Using this phytopathogen for our proof of concept was strategic as it enlights the power of our
tool and its utility.
For that, we interested ourselves in the economical impact of Dickeya solani
on the European crop losses. A Swiss case study (Dupuis et al., 2021) estimated these losses and
extrapolated them to Europe: 46 millions of Euros are lost each year in the potato sector due to
soft rot disease. Among these losses, the seed potato sector represents 32%, the consumption potato
sector (e.g. the potato that can be bought in groceries) represents 43% and the processing potato sector
represents 25%. The same study has also shown that the losses per hectare is 2.5 times higher for
the seed potato sector than the consumption potato sector, while the processing potato sector is
2.9-fold less affected than the consumption potato sector.
According to another study (Toth et al., 2011), the development of these bacteria and
the disease depends largely on climatic factors, such as temperature and humidity. Due to climatic
change, more than 25% of the blackleg diseases of potatoes have been attributed to Dickeya
species in the Netherlands (E. de Haan, NAK, Emmeloord, the Netherlands, personal communication with
the publication’s authors), Switzerland, Belgium (J. van Vaerenbergh, ILVO, Merelbeke, Belgium,
personal communication with the publication’s authors) and France (V. Hélias, FN3PT/RD3PT, INRA Le
Rheu, UMR lgepp, France, unpublished data). Today, Dickeya is the most frequent pathogen in
tubers. Moreover, since the appearance of Dickeya solani species, the losses caused by Dickeya
are increasing significantly, and the contaminated areas are spreading all over Europe, even in
Israel. These agricultural diseases can cause immense losses: as an example, in the Netherlands,
economic losses of more than 30 million euros have been caused by Dickeya every year
(Breukers, 2008).
In a more global vision, the spread of the epidemic of Dickeya solani is of great
concern because of two factors. First, potatoes are one of the most consumed crops world-wide.
Secondly, potato is the plant suffering the most losses in production due to pathogens (Savary
2019). The consequences of more losses can therefore be economically terrible and create a climate
of possible hunger in some parts of the world.
This case study shows that using FIAT LUX to fight epidemics, such as the one caused by Dickeya solani, represents a high solution to fight crop losses, food shortage, and economic losses. With our tool, this idea to fight Dickeya’s epedemic becomes more and more real. In fact our proof of concept worked, enlightening the applicability of our work in the real world! Because we always want to go further, we didn’t restrict our proof of concept to Dickeya but worked on other pathogens as well (see Proof of concept for more information), showing the extensibility and scalability of our project.
We wrote an awareness manual for farmers on plant health and
phytopathogens. This first edition is meant to be enlarged with more pathogens and with more
details acquired thanks to FIAT LUX. This is an example of how FIAT LUX could be
used to help farmers prevent crop losses.
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