Collaborations

Amoy

We collaborated with Amoy team by contributing to their newsletter. We have written about the selection process for iGEM in the No1 issue and participated in the questionnaire about the most useful software for us in No6 issue. We also published a description of our project in No3 issue, an update in issue No5 and abour our human practices in issue No7. Finally, we made a short assay about the situation of synthetic biology in Switzerland for the Special issue and about the ethical dilemmas of editing a human embryo for the No2 issue.

Darmstadt

We helped Darmstadt to evaluate their software.

EPFL

EPF Lausanne and ETH Zurich asked together people from the street about their opinions in synthetic biology

Stockholm

Starting from the very beginning of our iGEM summer we were in continuous contact with the team from Stockholm. We realized soon that our projects were going into a similar direction, both focusing on cancer detection in samples of bodily fluids, using engineered bacteria that will bind to cancer cells. In contrast to our project, Stockholm’s ABBBA is focussed on the detection of one specific type of cancer at a time. In several Skype meetings we exchanged our ideas about the best possible setup for single cell analysis and signal integration of the bacteria upon binding to a cancer cell. We provided the team from Stockholm with inputs on microfluidics as a setup for the final analysis and they helped us when we were struggling with the recovery of our gene fragments. But what seemed to be the best opportunity for combining our forces was the rather extensive experience in mammalian cell culture and bacterial-mammalian coculture our team had gathered in the course of our project. Since the team of Stockholm did not have the opportunity to test their system in such a real-life system, we decided to validate their system by combining their engineered E. coli cells with their actual targets: HER2-positive breast cancer cells.

Validation of ABBBA binding to cancer cells

We characterized the system of Stockholm regarding the binding of their bacteria to HER2-positive breast cancer cells. The affibody based system explored by the Stockholm iGEM team is designed to make engineered E. coli cells bind to cancer cells expressing the targeted surface marker. In the tested case of HER2-positive breast cancer, the team came up with three chimeric receptors, targeting different epitopes of the HER2 extracellular domain. We tested the binding of ABBBA E. coli expressing the respective constructs to HER2+ XXXX cells.

For better observation of colocalization of the bacteria and the mammalian cells we introduced a second plasmid carrying a constitutively expressed fluorophore (GFP?) into the bacterial strains that we received from Stockholm. We characterized the interaction of bacteria and mammalian cells after (link 2x) spheroblast and coculture protocols by (confocal) microscopy (and maybe FACS). The spheroblast protocol is necessary since the affibody construct is localized in the inner membrane of E. coli and has to be revealed by removal of the outer membrane.

Results

tbd

Characterization of lldR-operator promoter collection

In exchange, Stockholm agreed to characterize a big party of our synthetically designed lldR-operator promoters for us. They probed our engineered bacteria, containing a green fluorescent protein under the control of lldR along with constantly expressed lldR with or without lldP, for their reaction to different levels of L-Lactate, thereby greatly contributing to the characterization of our (link)promoter collection.