The team's accomplishments can be seen below. For more specific details on what each individual team member worked on this summer, see our Team Members page.
Seven different sgRNA candidates were successfully analyzed for CRISPR targeting efficiency, and their relative efficiencies were compared via indel formation rates.
Although the results are preliminary, the presence of a single adhered HEK293T cell in the puromycin-selected plate indicates the possibility of a functional CRISPR/Dual-Recombinase system. This result will require more time and sequencing to conclusively confirm, but we are extremely excited about the potential it holds.
Various chromatin modifying elements have been successfully cloned and sequence-confirmed and are ready for testing in eukaryotic cells. These parts, including a multiple cloning site optimized for dual-recombinase purposes, have been submitted to the registry for the convenience of future teams interested in recombinase.
The droplet formation module of our two-step microfluidics system has been prototyped and is in the testing phase, with plans to prototype the droplet electroporation module soon.
Furthermore, our Software Aggregating Research Assistant (SARA) is complete and functional, and available online for the use of future teams. This software will benefit future teams by compiling useful iGEM software for their own purposes.
None of these accomplishments would have been possible without the advice and support of our mentors who have led us along this journey.
Dr. Mayi Arcellana-Panlilio: Dr. Mayi Arcellana-Panlilio is the principal investigator for iGEM Calgary, and serves as a team mentor, organizer, and faculty liaison. Dr. Mayi, as we call her, is the glue which holds the Calgary iGEM team together year after year. Her unwavering support, enthusiasm and solid advice has been a pillar for this team to lean on. While she firmly believes in self directedness and allowing students to learn through trial and error, she was instrumental in troubleshooting and assay design. Perhaps most importantly, she supplied the team with endless smiles, love, and cookies.
Dr. Elke Monika Lohmeier-Vogel: Dr. Lohmeier-Vogel was the secondary PI for iGEM Calgary this year, and provided advice and support throughout the summer. Her knowledge of biochemistry was unparallelled during the frequent troubleshooting within the lab, and aided the team to develop solutions to their procedural pitfalls. Dr. Lohmeier-Vogel was a constant source of motivation and encouragement for the team members, supporting them both in and out of the lab.
Deirdre Lobb: Deirdre Lobb is the lab technician and manager for the iGEM Calgary lab space. She supported the team with material acquisition, providing countless chemicals and equipment throughout the summer. Deirdre also maintained safety and organization within the lab space, ensuring that experiments were conducted in a safe manner and that all materials were stored and handled properly.
Gene Integration Support
Dr. Aneal Khan: Dr. Khan's perspective as a medical geneticist, and the connections he made to people working in the area of cell and gene therapy, were invaluable in clarifying our vision of how Snip Equip Flip could be used in a clinical setting.
Dr. Ken Ito: Dr. Ito's cutting insight allowed us to place Snip Equip Flip within its competitive landscape. In particular, our decision to use wild type Cas9 rather than Cas9 nickase was a direct consequence of his comments.
Dr. Derrick Rancourt: Dr. Rancourt offered his usual, uniquely penetrating insight at several critical stages during the project's development.
Dr. Jonathon Lytton: Dr. Lytton discussed with us transfection methods for HEK293T cells and modification of cells.
Justin Simms: Justin discussed with us about the use of CRISPR and sgRNA design.
Iman Al Khatib: Iman discussed with us the use of RNPs and CRISPR, however her work involves mitochondrial DNA, so she introduced us to Tian Zhao.
Tian Zhao: Tian discussed with us HEK cell culturing techniques and various transfection methods in regards directly to CRISPR. He also attended our Faculty Talk.
Dr. Shirin Bonni: Dr. Bonni kindly gave us the HEK cells we worked with and also gave us valuable advice on transfections and transfection controls.
Gene Maintenance Support
Dr. Ray Wang: Dr. Ray Wang, who is a member at the Chen lab at the University of Calgary, provided us with Flp-In™ 293 T-REx cells. These cells will be used in future experiments and characterization of our Chromatin-Modifying Elements.
Dry Lab Support
Michaela Olsakova: Michaela tested our Software Aggregating Research Assistant (SARA) tool for usability and functionality. She also provided advice on wiki creation and feedback on the design and user-friendliness.
Patrick Wu: Patrick presented on wiki design at the North American Kick-Off and provided feedback on the presentation at aGEM.
Wm. Keith van der Meulen: Keith gave advice on effective wiki design and narrative creation.
Product Design/Modelling Support
Dr. Colin Dalton: Dr. Dalton has been instrumental in development of the microfluidics sub-project. He has offered us design advice, access to COMSOL Multiphysics, prototyping and measurement equipment, and access to his test labs. Aside from this, he has also provided us with excellent connections and advice on improving our presentation skills.
Dr. Ebenezer Owusu-Ansah: Dr. Owusu provided us with laser machining of the glass chips for all of our microfluidic devices. He had also provided us with insight into industrial level production and development projects.
Dr. Milad Azarmanesh: Early on in the summer we met with Dr. Azarmanesh to gain some insight into approaching design and experimental procedures for microfluidic devices. His comments molded our approach to developing this sub-project.
Human Practices Support
Dr. Walter Glannon: Dr. Glannon, a philosophy professor specializing in bioethics, met with members of the team to discuss the ethics of the project.
Dr. Ian Lewis: Dr. Lewis met with the team to discuss the potential applications of the project as well as providing direction regarding the narrative. He is the Health Solutions Chair in Translational Health in Metablomics at Alberta Innovates and is an assistant professor at the University of Calgary.
Dr. Philip Behman: The team met with Dr. Behman in August to discuss the social implications of our project, specifically from a Christian perspective. Dr. Behman is a spiritual care practitioner and member of the Spiritual Care Advisory Committee at Alberta Health Services.
Dr. Harjot Singh: Dr. Singh provided feedback from the Sikh perspective as part of the team's meeting with the Spiritual Care Advisory Committee in August. Dr. Singh is a family physician.
Fazel Rehman: Fazel is a community member of the Spiritual Care Advisory Committee at Alberta Health Services. He provided a Muslim perspective at our meeting with the committee in August.
Dr. Jaeun Macen: Dr. Macen is a spiritual health practitioner as part of the Spiritual Care Advisory Committee. We met with her and the rest of the committee in August, and she provided a Christian perspective to our discussion.
Kerri Kroeker: The team met with Kerri as part of our meeting with the Spiritual Care Advisory Committee. She is a spiritual health practitioner as part of the committee and provided a Christian perspective.
Tracey Staff: Tracey is a spiritual health practitioner as part of the Spiritual Care Advisory Committee. She represented a Christian perspective as part of our meeting with the committee in August.
Teresa Bond: The team met with Teresa, a spiritual health practitioner, as part of our meeting with the Spiritual Care Advisory Committee. Teresa provided a Christian perspective to our discussion with the committee.
Anna Majczak: Anna is a high school IB Biology teacher, the team met with her to discuss the implementation of an IBO curriculum addition and lesson plan.
Marija Susnjar: Marija is an IB Coordinator, the team met with her to discuss the implementation of an IBO curriculum addition and lesson plan.
Norman Hounjet: Norman is an IB Coordinator, the team met with him to discuss the implementation of an IBO curriculum addition and lesson plan.