Notre Dame Collegiate
We first came into contact with the Notre Dame Collegiate iGEM team at the aGEM competition in September, where we watched them present about their project "The RMS E. coli". They were working on creating E. coli that express an esterase capable of breaking down "fatbergs" that cause issues in wastewater treatment facilities. During this presentation, they indicated that they were interested in using spectrophotometry to gather quantitative data pertaining to enzyme kinetics under different reaction conditions. However, as a first year high school iGEM team, they had limited access to certain laboratory equipment.
We helped them design their spectrophotometry experiment and helped them run these assays using our spectrophotometer. They travelled from their neighbouring town of High River to our lab at the University of Calgary multiple times to attend a workshop hosted by our team's graphic design specialist and to perform the spectrophotometry experiments.
University of Lethbridge
Even before meeting at the North American iGEM Kickoff, the Calgary and University of Lethbridge iGEM team had hoped to build a proof of concept to integrate the strengths of their respective systems. It was a match made in biomolecular heaven: Calgary provided the gene editing machinery, while Lethbridge offered the delivery vector. By their forces combined, a scenario could be envisioned where Snip Equip Flip could be delivered in vivo, similar to a traditional viral vector, but while avoiding many of their intrinsic risks. After having brainstormed possibilities at the North American Kickoff for how the entire Snip Equip Flip system could be encapsulated in Lethbridge's PNCs, it was decided to first work towards validating that the encapsulated CRISPR component could be effectively delivered to mammalian cells in culture. For Snip Equip Flip to be fully implemented within a PNC-based delivery system, the donor DNA and integrating plasmid would also need to be co-encapsulated. Although several ideas were discussed for implementing such functionality, the development of those designs was decided to be beyond the scope of respective projects.
After leaving the Kickoff with renewed excitement for the collaboration, the teams went forward to plan the scope and details of their collaboration. It was then decided that P22 PNCs would be most suitable, and work proceeded in Lethbridge to prepare the Cas9-P22 scaffold fusion and P22 capsid vectors. An sgRNA generator, customized with Calgary's Cas9 target sequence, was also assembled. After meeting again at aGEM in late September, the Lethbridge team worked tirelessly to send purified proteins to Calgary. While the results are not ready for presentation on the wiki, both the Calgary and Lethbridge teams are hopeful that results will be prepared for presentation at the jamboree. Stay tuned!
As a fellow Canadian team, we hoped for an ongoing relationship with the Queens Canada iGEM team. Coincidentally, both members of the uCalgary team and the Queens team were also participating in a second research competition called the Canada Reduced Gravity Experiment (Can-RGX). Can-RGX is a nationwide competition wherein four collegiate teams were selected to prototype and test an experiment in micro-gravity through parabolic flight. Through Can-RGX, relationships were formed very early in the year between members of both teams, which continued to pay dividends for the remainder of the summer.
Cooperative planning and troubleshooting was performed throughout various meetings, both in person and over Skype. Topics such as the acquisition of funding, the ordering of complex DNA parts, and opportunities to take advantage of were shared and debated. The Queens Canada team alerted us to a promotion offered by Genscript which provided free DNA fragment synthesis, which proved integral to our project design. They also provided advice on re-designing DNA fragments which were too problematic to synthesize as single strands, aiding us in getting the ball rolling with our wet lab research. It was rewarding and refreshing to have a cooperative team with which we shared a running conversation and friendship, even for the simple ups and downs of undergraduate research.