Attributions

Project Design

The BeeT project consists of multiple parts, every student has their own part. We tried to integrate all the modelling with the wet lab. Therefore our project has three main categories, sensing the Varroa destructor, toxin production, and safety.

Wet lab

Lisa searched for the Bt toxin inside dead mites. Therefore we needed beehives from which to collect Varroa samples. Dead mites were collected from the beehives of the Duurzame Bij, a beekeeping organisation which tries to do beekeeping without pesticide use by breeding for Varroa-tolerant honeybees. They have some of the only beehives in the country with large populations of Varroa mites. Henk Kok, secretary from the Duurzame Bij, helped Lisa with the mite collection. We learned how to collect live Varroa mites and work with honeybees from Tjeerd Blacquière and Delphine Panziera. After the first few weeks, we were able to gather mites independently. It was a nice learning process and we learned how to avoid getting stung by bees. They also provided valuable input on honeybee research. Steven Aalvink gave a short instruction on the brightfield microscope. Sjef Boeren operated the Orbitrap for LC-MS/MS and provided instructions on how to prepare samples for proteomics. Kees van der Ark supervised Lisa's work.

Linea made a library of the toxin, she got some help in the lab from her supervisor Franklin, who explained how to work in the ML2 lab. Jaccoline made vesicles with advice from Vera Silva. Jaccoline also worked together with Linea. She learnt about DLS (dynamic light scattering) from RG. Fokkink who also explained how to analyse the data. With this information Jaccoline could analyse her results. Carina made the Varroa sensing system based on sensing mite poop and vitamin B12.

Thomas made a quorum sensing system and a subpopulation system. Thomas was supervised by Franklin during the building of this system and testing it in the lab. Daniel Gerngross made the RBS library with RedLibs algorithm for Thomas. Thomas tested this RBS library in the lab himself. Tjakko Abbee from the Food Microbiology group at Wageningen UR gave him permission to use the FACS for free. Marcel Tempelaars from this department helped Thomas to do flow cytometry, he explained to Thomas how it worked and did the measurement together with him. Before that Thomas tried to do FACS in Leiden with help of Paul van Heusden but unfortunately this was unsuccessful.

Belwina used Cas9 to improve a biocontainment kill switch. To design the genetic circuit, she was helped by her supervisor and her second supervisor Franklin. Her supervisor Alex also helped out with lab work by handling the FPLC machine. Ioannis Mougiakos recommended the Expresso system for getting Cas9 expressed in the right amount for doing in vitro Cas9 assays. Prarthana Mohan Raju helped by advising on how to perform the assays and cloning and expressing the gRNAs.

Marijn made the light kill switch in the lab and he did also light measurements in the beehives. Marijn got some help from his supervisor Marta. Tianhe took care of making a toggle switch and combining the light kill switch and sensing system together. Tianhe got help from his supervisor Stamatios

Modelling

Mario modelled the light kill switch which, he got advice of how to do dynamic modeling by his supervisors Emma and Rob. Remco modelled the interaction between the BeeT, Varroa, and Bees to investigate how BeeT would spread through the hive and affect the mite and bee population. His teammate Ronald explained to Remco how to use R and also helped him with bash related problems he encountered in the process. His supervisors Rob and Emma gave him advice.

Angelina modelled the quorum sensing subpopulation system. Her teammates Remco and Ronald helped her with some of the problems. Her supervisor Rob gave her advice on how to make the model and Ronald worked on the metabolic modelling and an in vitro toxicity assay, he got help from his teammate Thomas when he went into the lab. He was supervised by Maarten Reijnders.

Human Practice

Carina is our Captain Human Practice, Carina forms the Human Practice team with Linea and Belwina. As one of the first things they did was get in contact with Beekeepers association in Wageningen (NBV). Together with them we tried to think of a solution for the dying bees. We have a collaboration with Synenergene where an application scenario was generated. A movie about synthetic biology has been made for the RIVM. We also had a collaboration with the Design Academy Eindhoven, where Thieu Custers made our product design.

Wiki Construction

Lisa is the Wiki Captain, together with Ronald, Remco and Tianhe the design of the wiki was made. We got advice from the Rik van Rosmalen on making the Wiki. Lisa made all the illustrations herself and together with the guys did most of the programming work. Every team member worked hard to get his/her personal pages on the wiki.

Finances

With Angelina as our Treasurer, the finances were managed together with Jaccoline for the crowdfunding. For the crowdfunding we got help from Arianne van Ballegooij, she told us how we could sell ourselves the best. Mario looked for the plane tickets and accommodation. As a team we searched for sponsors and got advice from our supervisors, specialists at the university and previous iGEM team members. We searched all together for sponsors, we asked the previous Wageningen UR team which companies they would recommend us to approach.

Other Team Roles

With Thomas as our Team Captain the whole team functioned well. Of course he had much help from his secretary Belwina. Linea is Captain Facebook, nothing is more important these days than presenting yourself on social media. Marijn is our lab safety manager who took care of keeping the lab safe and clean.

Presentation

During the iGEM project, we had multiple presentations. We organised a presentation open to all people, staff and students, at Wageningen UR. The help we got with the preparations was from the supervisors. They sat together with us and helped us with our presentation skills. They also commented on the presentation slides, told us what was clear and what was not. Impulse provided us with a venue and sponsored lunch during our public presentation.

Supervisors

Dr. Christian Fleck, prof.dr. John van der Oost, dr. Robert Smith, Kees van der Ark, Maarten Reijnders, Stamatios Damalas, dr. Marta Vazquez Vilar, Emma Keizer, Alex Kruis, and Franklin Luzia de Nobrega. Our supervisors!

Advisors

Bob Mulder gave us advice on how to approach beekeepers.