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 the Varroa. Therefore we needed beehives from which to collect Varroa samples. We learned how to collect live Varroa mites 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. 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. Linea made a library of the toxin, she got some help in the lab from her supervisor Franlin, 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 Vitamine B12.
Thomas made a quorum sensing system and a subpopulation system. Thomas got help with building his system and testing in the lab by his supervisor Franklin. Daniel Gemgross made the RBS library with RedLibs algoritm for Thomas. Thomas tested this RBS library in the lab himself. Prof. Tjakko Abbee form the Food Microbiology group at the Wageningen University 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 Assis. Prof. Paul van Heusden but unfortunately this was unsuccessful. Belwina used Cas9 to improve a biocontainment killswitch. To design the genetic circuit, she was helped by her supervisor and her second supervisor (Franklin Nobrega de Luzia). Her supervisor also helped out with labwork 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, Tianhe took care of making a toggle switch and combining the light kill switch and sensing system together.

Modelling

Mario modelled the light kill switch which, he got advice of how to do dynamic modeling by his supervisors Emma and Rob. Marijn was making the light killswitch in the lab. Remco modelled the interaction between the Varroa and Bees and the spread of toxin through the hive. His teammate Ronald explain to Remco how to use R and also helped him with bash related problems. His supervisors Rob and Emma gave him advice. Angelina modelled the quorum sensing subpopulation system with lot of bugs she created herself when writing the code for the system. Her teammates Remco and Ronald helped her fix the problems. Her supervisor Rob gave her advice on how to make the model and Ronald worked on the metabolic modelling and a in vitro toxicity assay, he got help from his teammate Thomas when he went into the lab.

Safety

As for the safety we want to implement a light kill switch in our end product. Also the Cas9 should prevent the bacteria from spreading through the environment.

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. 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.

Lisa is the Wiki Captain, together with Ronald, Remco and Tianhe the design of the wiki was made. We got advice form theiGEM Wageningen_UR 2014 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 there own 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 Ballengooi, 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 team which companies we could ask and what was the best way.

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 at social media. Marijn is our lab safety manager who took care of keeping the lab save and clean.

During the preparations for the iGEM project, we had multiple presentations. For example for the University. 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.

Supervisors

Christian Fleck, John van der Oost, Robert Smith, Kees van der Ark, Maarten Reijnders, Stamatios Damalas, Martha Vazquez Vilar , Emma Keizer, Alex Kruis, Franklin Nobrega de Luzia.

Bob Mulder