Based on feedback from experts in various fields, we modified our construct and system’s designs. Initially, we planned to deliver antimicrobial peptides to cyanobacteria, but after receiving critiques about the need for increased target specificity, we switched to a CRISPR Cas13a system, as it allows us to target a gene that is unique to Microcystis aeruginosa cyanobacteria. To ensure the safety of our treatment, once we obtain preliminary data on the functionality of our system, we will perform various rigorous tests that were advised to us. We have also discussed with experts on the use of mesocosm and experimental lake studies, essential additions incorporated in our experimental flow chart. The use of mesocosm and experimental lake studies provide an imperative expansion to regular laboratory bench experiments, which are limited in their ability to simulate aquatic environmental conditions. Finally, molecular modelling will allow us to optimize our system’s design, as well as improve both binding affinity and specificity. The advice provided through interviews of various experts have helped shape and improve our project this year.
Biosecurity and Biosafety
Members attended a dual use research of concern (DURC) workshop organized by the Alberta RNA Research and Training Institute at the University of Lethbridge, in collaboration with SynBio Canada. From the knowledge gained during this workshop we discussed potential biosecurity risks surrounding our proposed treatment method. Regarding our proposed delivery, suppression of cyanobacteria growth, and cyanotoxin neutralization, we considered potential malicious misapplications. This includes the replacement of the target sequence in the crRNA of the CRISPR Cas13a system to target other organisms that are essential to the environment. However, there are chemical treatment alternatives that are cheaper to make or purchase that cause more harm to ecosystems.Therefore, engineering a synthetic biology treatment is a promising method to solving the issue of cyanobacteria.
As for biosafety, the CRISPR Cas13a system utilizes a crRNA that interacts with cyanobacteria mRNA. As a result, it does not interact with DNA, circumventing any danger of changing the genomes of organisms in the environment. Furthermore, if our treatment results in eradication of cyanobacteria from the lake ecosystems, this could cause a bottom up ecological cascade. To address these issues, we plan to test the specificity of the target sequence and the efficiency of our project first within laboratory conditions and afterwards within mesocosms and later experimental lake facilities.
Potential Customer/real world implications
We have determined three possible demographics that would be interesting in the use of our system; government, agriculture, and the water activity enthusiasts.
As the federal and provincial governments heavily regulate treatment applied to bodies of water, this demographic will play a big role. To discuss what steps we would need to take and what these policies may look like, we plan to get into contact with individuals involved in regulation and policy making regarding synthetic biology applications to bodies of water.
The second demographic we have identified includes the agriculture sector. Albertan farmers often rely on man-made bodies of water that can be prone to cyanobacteria blooms, such as farm dugouts. While there are more treatment options applicable to man-made bodies of water, they still have their limitations. For example, many farmers choose to raise fish in their dugouts or have animals such as cattle that have access to the water. Chemicals such as copper sulphate treatments are toxic to beneficial aquatic organisms as well as animals. However, because of these off-target effects, many products using copper sulfate are no longer used or legally sold in Canada.
The third demographic that may be interested in our proposed solution is the general public. Various recreational aquatic activities are interrupted each summer when cyanobacteria blooms form. Some examples of activities interrupted by cyanobacteria blooms include:
Fishing: toxins can persist in fish from lakes with cyanobacteria blooms
Swimming and other water sports
Boating
Pet owners
Government and Regulations
All herbicides and pesticides are regulated by the Canada Pest Management Regulatory Agency and must be registered for use. The Environmental Protection and Enhancement Act (EPEA) covers pesticide regulations and fertilizer use, storm water drainage and municipal wastewater treatment. Under Canadian and Alberta laws, chemical treatments in natural bodies of water are heavily restricted. Most treatment options are only approved in man-made bodies of water such as dugouts. Furthermore, these laws do not cover synthetic biology systems such as CRISPR Cas13a.
Entrepreneurship
This year many of our team members have had the wonderful opportunity to take part in an independent study-like course that is being run by Agility, a campus-wide program that is 100% donor-funded. Agility supports students of all disciplines interested in entrepreneurship and innovation, and provides instruction and tools to students who wish to learn more about those topics. The manager of this program, Brandy Old, is dedicated to coaching student entrepreneurs and encouraging the thought processes and mindset that will help future entrepreneurs to thrive in the modern market. Although the team is not finished with this course, one of the skills in particular, market discovery, will be a great help later in the project’s lifespan, when we are looking into acquiring more funding, attracting sponsors, and trying to enter collaborations with other organizations and companies.
Awards and Fundraising
This year we have been able to secure a considerable amount of funding thanks to these sponsors. MindFuel, a nonprofit organization that supports STEM programs in Alberta, provided us with a grant after our presentation at their Tech Futures Challenge, where we were awarded second place. Genome Alberta, an organization that promotes the advancement of genomics-relevant technologies and research in our province, has also supported us financially. We have also received funding as part of an ISEE grant or Institutional Support for Entrepreneurship Education from Alberta Innovates. Finally, the University of Lethbridge provides in-kind support to our team by giving us access to laboratory space and mentorship by a faculty member and two graduate assistant students.
Collaborations
Lethbridge High School iGEM team
Wiki Workshop
Members of our team took part in a Wiki-Workshop, which included an overview and tutorial for the Lethbridge High School team's iGEM members on how to set-up and edit a wiki using HTML
Construct Help and Improvement
High River and Canmore HS iGEM teams
Modelling
Construct Design
Outreach and Volunteering
Recital
Mark Lea, one of our student leaders, performed at a recital organized by the Lethbridge High School Team
Coulee-Cleanup
This year’s team participated in a coulee clean-up in an effort to maintain and promote the health of our natural community with the supplies donated from The Helen Schuler Nature Centre. Our team members removed waste items from our coulees, as well as items that may pose a hazard to the wildlife found in the coulees of Lethbridge.
HYRS
Our team put together a presentation on 'How to do a scientific poster presentation' for the HYRS (Highschool Youth Researcher Summer) program. The presentation included information about our project and iGEM in general.
