- Included a plasmid addiction system, and regulation for CRISPR/Cas9, based on Dr. Wanyin Deng's concerns.
- Changed our target gene from VirB6 to VirD2, and used RP1 as our plasmid backbone, based on Dr. Stanton Gelvin's suggestions.
- Arranged a meeting with representatives of the Public Health Agency of Canada to discuss public biosafety implications of aGROW, after discussion with Dr. Guus Bakkeren and Dr. Linda Delli Santi.
- Created our protoype with respect to grafting remediation instead of soil remediation, based on advice from Dr. Xin Li and Dr. Wanyin Deng.
- Made science communication and education among all age groups our primary human practices mission, after our interview with Dr. Santa Ono.
- Developed camps, workshops, presentations, videos, and founded a club to help dispel misconceptions about genetic engineering, after our discussion with Dr. Linda Delli Santi.
- Published educational materials for others to use to help educate and inspire the next generation of scientists and engineers.
Integrated Lab Work
Our conversations with local experts from industry and academia helped us identify areas where our project could be improved.
Dr. Deng recommended we include a positive regulation mechanism to ensure our engineered plasmid was not lost through plasmid curing. To address this, we included a plasmid addiction system into our design and investigated both the hok/sok addiction system and a toxin/antitoxin system native to Agrobacterium tumefaciens, ietA and ietS. Additionally, Dr. Deng indicated that constitutive expression of CRISPR/Cas9 could reduce the efficacy of our system. To avoid this, we investigated opine regulated promoters to regulate CRISPR/Cas9 expression as a similar system is already present in A. tumefaciens to control conjugation
Dr. Gelvin suggested that we change our targeted gene from VirB6 to VirD2,as VirD2 is more conserved throughout Agrobacterium and Rhizobium populations. From this conversation, we changed the genes inputted into our model and altered the guide RNA design to incorporate this information. Initially, we also wanted to upregulate conjugation by constitutively expressing the conjugative pathway. With Dr. Gelvin’s recommendation, we investigated the IncP set of plasmids. After a thorough investigation, our team decided to use RP1 as our plasmid backbone for the final prototype.
Based on the safety concerns of Dr. Delli Santi and Dr. Bakkeren, we wanted to ensure that our system was safe for long-term commercial applications. Throughout project design and planning, we were careful to thoroughly consider the potential outcomes of the parts included in our prototype. Additionally, we have contacted and arranged a meeting with representatives of the Public Health Agency of Canada at the iGEM competition to discuss the safety implications of our project.
Several of our industry experts, including Dr. Li and Dr. Deng indicated that for industrial crops, soil contamination was not as much of an issue as grafting contamination. While this concern would only be relevant for a final stage product, we theorized about how we could apply our prototype to grafted plants, rather than the soil environment. This distinction allowed us to gain credibility with some industry professionals and guided our long-term development strategy.
Integrated Community Involvement
In our interview with UBC President Santa Ono, he emphasized the importance of science education and communication, especially when dealing with topics like synthetic biology. We decided to make science education and communication the primary focus for our human practices program. This focus led us to develop and teach a kids camp curriculum for several weeks throughout the summer. We wanted to be able to get young kids interested and engaged in STEM, and help give rise to the future generation of engineers and scientists. A complete description of our kids STEM summer camp can be found under Engagement and Education.
In our conversation with Linda Delli Santi, the executive director of the BC Greenhouse Growers’ Association, the importance of the public conception of GMOs was mentioned. In order for our project to be successful in its commercial use, she indicated that public understanding and knowledge of genetically modified organisms needed to be improved. This led us to develop a workshop curriculum to educate middle school and high school students about synthetic biology and clarify any misconceptions they might have. To reach a university level audience, several of our team members came together to form a brand new club on campus - the AMS Synthetic Biology Club. The goal of this club is to spread knowledge and awareness about genetic engineering with our peers. We believe that creating a knowledgeable foundation about genetic engineering will allow students to make smart, informed opinions on GMOs in their future. We also produced videos about iGEM, our project, and the work done with synthetic biology, and shared them on our Facebook page. To read more about our genetic engineering workshops, Synthetic Biology Club, and Facebook videos, visit the Engagement and Education page.
All of the activities and games we developed are published on our wiki for other future iGEM teams to use when developing workshops for students. A complete curriculum of our kids camp activities is also available, which include all of the projects we did to help develop an interest in science and engineering for younger kids. We hope that these materials will be able to go to good use - inspiring an interest STEM fields in students, and building a generation of individuals educated about GMOs. Check out our games, activities, and protocols under the Engagement and Education page.