On January the 5th, we had our first meeting with the advisors and the students of the former team. We were introduced to the competition, and last year’s team explained to us the main roles. It was decided that we would have a regular meeting every Thursday, lasting about one hour and a half during lunchtime. Throughout the week, we discussed and decided on the ideal roles for each member of the team within the organization and reviewed the projects we worked on for our applications.

Roles have been assigned as follows: Organization (Cléa), Communication (Bayane), Experimentation (Pakindame), Wiki (Koloina), Human Practice (Héloïse), Finances (Mathieu), Modeling (Jeanne) and Entrepreneurship (Marie).

We also discussed three topics we found interesting to develop:

• - biological glue
• - biological light
• - oil spill treatment

From these discussions, some new ideas emerged, such as using biological light to illuminate tissue during an operation to assist the surgeon.

Weekly zoom meeting with our advisors.

This week, we continued brainstorming on the three topics we discussed the week before. Due to exams, it was challenging to find time to explore new subjects. We started creating slideshows to present our ideas to our supervisors.

Slideshows to present our ideas each week

We started looking for new ideas. We abandoned the idea about a biological glue because it had already been done.

We started meeting more often every week, including weekends. We created a poster for the open day of INSA.

During our regular meeting, each advisor was assigned a role to help us and answer our questions. We continued brainstorming about bioluminescence, and the idea of using cell-free expression in liposomes was mentioned. The goal would be to induce light when the liposome binds to cancer cells, illuminating the operation zone. We also thought about directly delivering anticancer drugs to the tumor site instead of illuminating it. As for oil spill treatment, we reached a deadlock and decided not to continue research on it. Additionally, we discussed a method of detecting endometriosis.

During the regular meeting, new ideas were discussed. We also continued our discussion about using liposomes to fight cancer cells.

The first team building event was organized, where we enjoyed tartiflette, a French cooking specialty, and played board games together.

Tartiflette savoring

On February 11th, we participated in the school's opening day to present the competition and the projects of previous teams to high school students.

INSA's Opening day

We also met with Thomas Esquerré, an anesthetist at the University Hospital of Toulouse, to discuss the interest of our idea, which is using liposomes to fight cancer cells. He explained to us the methods currently used during intraoperative procedures. You can find more information about this interview on our IHP page.

Meeting with Thomas Esquerré

We came close to choose the liposome topic but decided to keep the possibility of finding new ideas for one last week. We discussed the molecular aspect of the project and divided it into three parts to facilitate the research:

• - the binding to cancer cells
• - the signal transduction
• - the production of anticancer drugs

The first meeting to begin organizing a French meet-up in Toulouse was held. The BioEco graduate school is partnering in this project. We got in touch with iGEM Nantes to explore the possibility of collaborating to organize this event.

This week marks the final selection of our topic on the use of liposomes to kill cancer cells! We continued our research on the molecular aspect of our project, as well as on current treatments for cancer and their limitations.

The first meeting with iGEM Nantes took place, and we decided to collaborate between our teams to organize this mini-jamboree in Toulouse.

Meeting with iGEM Nantes

We met with Jean-Pascal Capp, a teacher-researcher at Toulouse Biotechnology Institute who used to work in the field of cancer. He confirmed some of our choices regarding our system and provided us with new leads for anticancer drugs.

Meeting with Jean-Pascal Capp

We listed all the different molecular possibilities for each part of our project and created diagrams to illustrate our ideas.

We met with Dr Ilfad Blazevic, an oncologist in IUCT-Oncopole at Toulouse to have his expertise on our project. During the meeting, we also gained more insights into the various types of treatments for cancer and their limitations.
Please, consult our IHP page to get more information regarding this interview.

Meeting with Ilfad Blazevic

We also started reflecting on a project name.

A second team building event was organized with our advisors, and everyone showcased their cooking skills!

We started discussing the experiments we would need to conduct this summer for our proof of concept.

This week, we had a meeting with iGEM Nantes and Eur BioEco to finalize the date of the meetup! It has been scheduled for the 22^nd and 23^rd of June.

Meeting with iGEM Nantes and Eur BioEco

We also found the name for our project: CALIPSO which stands for Cell-targeting Advanced LIPosomes for Selective Oncotherapy.

This week marked the start of our regular meetings with the iGEM Nantes team to organize the meetup. We discussed the event's outline, including the welcoming speech, conference, and mini-jamboree.

Meeting with iGEM Nantes

We also decided to focus our human practice approach on patients to better understand their living conditions.

Additionally, we began discussing the genetic constructions we would need for our experiments.

We had a meeting with Elvire Prochilo, an entrepreneurship and management consultant, who provided valuable guidance on conducting our entrepreneurship project. She gave us advice on the business plan and other aspects.

In addition, we created a draft for our logo.

We started familiarizing ourselves with Benchling and searched for the protocols.

Additionally, we designed an Instagram post for the meetup and reached out to the French teams to inform them about the event.

This week, we finalized our modeling strategy, aiming to focus on both the structural and functional aspects of the project.
Visit our modeling overview page to find out more!

We also had a team building meal together at a restaurant!

Burger savouring

This week marks the official beginning of the iGEM adventure with the team’s registration for the competition!

It has been an intense week of work as we started to design our biobricks on Benchling to ensure we can order them on time. We also prepared a list of everything we would need for the experimental part of our project and secured sponsorship from GeneFrontier to provide us with the PURE system solution.

During this month, we secured sponsorship from the Toulouse Biotechnology Institute (TBI) and Toulouse White Biotechnology (TWB)! We also initiated contact with various companies in France, although we received several negative responses.

Logos of our new sponsors

We started placing orders for certain products, such as lipids, and continued the process of designing our biobricks. Additionally, we created diagrams to visualize all the steps required to demonstrate the functionality of our system.

Perseo Pharma announced its sponsorship for our project!

We met Philippe Urban, an expert in protein biochemistry, who assisted us in using AlphaFold to determine the structure of the T7 split.

We began to prepare for Exposciences, a scientific event aimed at popularizing science to young people aged 5 to 25 years old. For our participation, we created two posters: one explaining synthetic biology and another detailing different types of treatments. We also prepared activities to explain the fundamentals of synthetic biology, such as transcription and translation. See explanations of the game in our Education and Communication page

We had a meeting with a teacher-researcher from our school, Jean-Pascal Capp, to gather more information about cell culture and general safety protocols. We discussed which cell lines would be suitable for our experiments and how to handle them safely.

Our collaboration with the Catalyseur of Toulouse, a business incubator, has started! We had a meeting with Colette Schenker, who is in charge of the support program for entrepreneurs and start-ups. She provided us with valuable advice to improve our presentation skills.

We also ordered our gBlocks for our parts from IDT!

This was the final week of our classes.

On Saturday, we took part in Exposciences. It was a great opportunity to learn how to present our project to the general public. We also recorded an episode of the podcast "Pose ta science" with Radio Francas. You can find more information about this event in our Education and Communication page

Photos taken during Exposciences

This week marked the beginning of our internship! It was a busy week as we received materials for our experiments, organized our lab and underwent security training for the use of autoclaves.

First day of lab

On Tuesday, we pitched our project during the annual Comité of the Catalyseur to other entrepreneurs.

Oral presentation at the Catalyseur

We also spent the week calling companies and public organizations to raise funds. Unfortunately, many of our emails went unanswered.

To be more efficient in lab, we divided the different responsibilities as follows:

• - Cloning: Pakindame
• - Pure system: Mathieu
• - Liposome preparation: Bayane
• - Cell culture: Jeanne
• - Expression and purification of DhdR: Cléa
• - Expression and purification of anti-Her2 nanobodies: Héloïse
• - orders: Koloina/Marie

We started the cloning process for one construction.

Progression of our constructions

The Oncopole celebrated its 100th anniversary during which we discussed our project with professionals and made contacts for our IHP.

We finally selected our project’s logo and graphical charter!

The cloning process continued.

Progression of our constructions

This was a really intense week as we had to handle the last-minute details of the meetup. The meetup took place from Thursday to Friday and everything went well. It was a great opportunity to exchange with other iGEM teams and learn more about their projects. Visit ourcollaborations page to learn about the course of this event!

As researching sponsors proved to be very challenging, we launched our crowdfunding campaign to raise additional funds (see our Acknowledgment page).

This month, the Paul Sabatier University and Lesaffre announced their sponsorship of our project.

We also shared a press release to present our project to the public.

Experimentally, our clonings were successful for DhdR, sfgfp and typh constructions. However, we encountered problems with the T7 constructions.

Progression of our constructions

Experimentally, we initiated the production and purification process for DhdR. Additionally, we discussed and tested various strategies for cloning the T7 constructs.

We pitched our project at the Summer Tech day, an event that brings together entrepreneurs and investors.

Pitch at Summer Tech day

Finally, we organized several meetings with healthcare personnel as part of our human practice approach. For instance, we met with two advanced practice nurses, Kristell Thomas and Alexandra Lambert, as well as Anne Gauci, a healthcare manager. Please consult our IHP page to learn more about these interviews.

We wrote the storyboard of our promotion video and produced drawings for the animation.

We initiated experiments using the PURE system to produce thymidine phosphorylase, anti-Her2 nanobodies and sfGFP in bulk. The results were conclusive only for sfGFP.

This week, we continued to meet with healthcare staff. We had discussions with Muriel Richl, a dietician at Oncopole, and with Aurélie Robello, who is responsible for the therapeutic workshops. This marks the beginning of organizing our own workshop on Synthetic Biology with patients.

We began the production and purification process for anti-Her2 nanobodies while the cloning for the T7 constructs remained ongoing. We restarted the production and purification of DhdR, and it was successful! See our results here.

We also presented our project at the BioSynSys Symposium held on the campus of INSA Toulouse. This provided us with an opportunity to receive feedback from the scientific community regarding our project.

Finally, we learned more about intellectual property by discussing it with Philippe Lucas from the INPI (National Institute of Industrial Property).

We attempted, for the first time, to measure the repression of DhdR on the expression of sfGFP in the PURE system. We also determined the concentration of the produced nanobodies.

In addition, we booked the train and the accommodation for our trip to the Jamboree.

Finally, we began filming our project promotion video!

This week was different because we didn’t have access to our lab. As a result, we spent the week writing our wiki and strategizing for entrepreneurship. We enjoyed “sweet” breaks to recharge our energy.

We also recorded a podcast with "Good Morning Toulouse" radio, where we discussed our project and the competition.

We finally received our t-shirts!

A perfect timing since a journalist came to our laboratory to write a report about us. Find the link to the article on our education and communication page.

Experimentally, we restarted the cloning process for the T7 constructs using the pET21a (+) instead of the p05-79 and p05-39 plasmids. We also conducted tests using various concentrations of DhdR on the G-block containing the GFP.

Experimentally, we finally succeeded in cloning the Pertuzumab antibody linked to the Nterm of the split T7. We attempted to produce this complex in the Puresystem.

Simultaneously, we were in contact with the student responsible for creating the video animation, Oscar Baron. Additionally, we also received the first draft of the video's musical composition from Léo Vendeville.

We shot several scenes for our video including those captured with a drone.

Many meetings were held to discuss our entrepreneurship and HP strategies, as many people's holidays were coming to an end. We had the opportunity to make progress on our educational workshop.

We also had our first meeting with the Patras-Medicine iGEM team to discuss how we could collaborate together. It was so nice meeting another team and presenting our respective project!

Experimentally, we successfully prepared a lipid film hydration for the first time using lipid-coated beads. We also cloned the SP6 promoter on the rapamycin construction. Finally, we conducted tests to detect the pro-drug and drug using HPLC.

For our promotional video, we recorded the voiceover and completed the editing.

For the first time, we tried to visualize liposomes expressing sfGFP by fluorescence microscopy. We also succeeded in expressing the anti-rapamycin T7 RNA split polymerase in the PURE system. We planned the remaining experiments to be conducted by the end of the internship by creating diagrams.

Regarding our educational workshop, it was too difficult to obtain the necessary authorizations to organize it in the hospital location. We had to make urgent phone calls to find an alternative venue for it. In parallel, we completed the cards for our game.

Experimentally, we conducted toxicity tests with 5-FU and Tegafur on cancer cells. We also assessed the impact of Tegafur on the synthesis of sfGFP in the PURE system. Additionally, we attempted to demonstrate the activity of the anti-rapamycin T7 split in the PURE system.

We started creating the storyboard for our presentation video and put a lot of effort into writing the content for our wiki.

Finally, we tested our game with some friends to further refine the concepts and format. You can find more information on the game in our education and communication page.

We visualized the anchoring of liposomes decorated with anti-HER2 nanobodies to Caco-2 cancer cells! To achieve this, we utilized the facilities at LITC (Light Imaging Toulouse CBI) at the CBI (Centre de Biologie Intégrative).

We also decided to compete for the best measurement, best model and best entrepreneurship prizes.

Additionally, we conducted our synthetic biology workshop at a junior high school, where the students were receptive to our explanations and appreciated the experiments!

Finally, regarding entrepreneurship, we made the sizing of our prospective lab: consult our Implementation page to explore our 3D lab!

This week concluded our time in the lab, indicating the end of our internship. As a result, we carried out our final experiments, which included toxicity tests of 5-FU on cancer cells and producing liposomes. Notably, some of these liposomes demonstrated the ability to express the sfgfp gene after incubation in a culture medium with cancer cells!

Last day of lab

Thanks to our collaboration with Etincelles association, we were able to meet with patients and host our workshop called “Biology to better understand one’s illness”. This workshop proved to be invaluable for both the patients and ourselves.

Finally, we put a lot of effort into redacting our wiki as the wiki freeze was getting closer.

This week marks the beginning of the school year, which meant we had less time to dedicate to our project. It was a demanding week as we needed to finish our wiki and complete the registry.

In parallel, we filmed our presentation video.

We checked one last time every detail of the wiki and the registry to be ready for the wiki freeze and started to prepare the final presentation. Thank you for reading the entire Notebook. As a reward, here is one of our most recent photos of us enjoying a meal (for a change)!