Q&A with the CAPS Kansas iGEM Team

Twist Bioscience is proud to be one of the sponsors of the Blue Valley Schools CAPS Kansas iGEM team. We took some time to catch up with them as they were finishing their preparation for the 2016 iGEM Giant Jamboree.
Twist Bioscience: Who are the Blue Valley Schools CAPS Kansas iGEM Team?
Blue Valley Schools CAPS Kansas iGEM Team: The Center for Advanced Professional Studies (CAPS) high school iGEM team is located in the heartland of the United States, in the state of Kansas. The team includes Steven Blair, Hanna Bradford, Grace Brunner, Hayden Dahm, Archiata Goyal, Joshua Jeng, Lucy Li, Nithin Saripalli, Anna Song, Archana Sundar, and Emma Van Lieshout and is coached by Eric Kessler and Joe Whalen, instructors at CAPS. Team members are sophomore through senior students at one of the five high schools in the Blue Valley School District. Some of the team members are enrolled in bioscience courses at CAPS while others participate in the team through our after school bioclub. More information can be found on our iGEM team website.
Twist: How did you come to compete in iGEM?
CAPS Kansas: One of our CAPS bioscience instructors, Eric Kessler, learned about synthetic biology and iGEM through interactions with Dr. Todd Eckdahl at Missouri Western University and Dr. Malcolm Campbell at Davidson University, who used to maintain a joint iGEM team. In the summer of 2011 Eric observed the first high school iGEM Jamboree held in Greenfield, Indiana. Returning quite motivated from the event, CAPS participated in subsequent high school iGEM Jamborees held in Greenfield and in Boston from 2012-2014. In 2012 we tied for Best Poster, and in 2013 we happily accepted the iGEMers Award. We are proud to be back competing at iGEM this year! 
Twist: What is your project all about this year?
CAPS Kansas: Ever since our first brainstorming meeting in the fall of 2015, team members have been interested in tackling the growing problem of antibiotic resistance. Nithin in particular was persistent in motivating the team to consider ways we could use the emerging CRISPR/Cas9 technology. Early this summer, we hypothesized that the AcrAB-TolC efflux pump, important in the expulsion of antibiotics from resistant cells, might be a good target for such technology.
In our initial experiments, we designed guide RNAs to target promoters or coding regions in the genes for AcrA, AcrB, and TolC, and cloned oligos encoding these RNAs into a plasmid designed by the Stanford-Brown 2013 iGEM team. Upon the recruitment of dCas9 to the sites targeted by these guide RNAs, transcription by RNA polymerase should be inhibited. We expect that expressing these CRISPR/dCas9 systems will result in E. coli that are more susceptible to antibiotics, as measured by the Kirby-Bauer test. The Kirby-Bauer test is performed by placing a small antibiotic-containing disk onto a freshly-spread plate of bacteria and observing the subsequent growth patterns.  If bacteria are susceptible to a specific antibiotic, there will be a “zone of clearing” around the antibiotic disk where there will be no bacterial growth.  By contrast, if bacteria are resistant to a specific antibiotic, there will be bacterial growth on the plate surrounding the disk.
“We are inspired by the idea of exploring novel synthetic biology techniques for confronting the problem of antibiotic resistance without the need for the development of new antibiotics.”
Twist: What inspired your project?
CAPS Kansas: The development of antibiotics was one of the most significant human accomplishments in the past century. The recent reduction in the rate of new antibiotic discovery coupled with an increase in the evolution of antibiotic resistant bacteria is threatening the legacy of this achievement. We are inspired by the idea of exploring novel synthetic biology techniques for confronting the problem of antibiotic resistance without the need for the development of new antibiotics. 
Twist: Are there any moments that are particularly memorable from your project?
CAPS Kansas: To date, the most memorable moment occurred when we didn’t observe any bacterial colonies in our Petri-dishes after completing our initial transformation of the Stanford-Brown 2013 part from the iGEM 2016 distribution. We were quite depressed and began troubleshooting to identify our possible mistakes. After allowing our liquid cultures to grow another day and plating them again, we discovered that our transformation had actually been successful!
Twist: Are there any moments that stick out as particularly challenging over the course of your project?  How did you overcome them?
CAPS Kansas: We have confronted a number of challenges during this iGEM season. We had difficulty coming up with a specific means of applying CRISPR to the problem of antibiotic resistance, maintaining team continuity during the summer when everyone had different schedules, raising money to fund our iGEM registration and our trip Boston, and finding enough time to get things done once school started—to name just a few. We haven’t overcome these challenges entirely but through the shared leadership of our team members, the guidance of our mentors, and the excitement of engaging in cutting-edge biology, we have continued to progress one step at a time.
Twist: If you had another six months, where would you take your project?
CAPS Kansas: Once we are successful in testing our current guide RNAs within E. coli, we would like to explore methods for delivering for our CRISPR/dCas9 system to other bacteria. This would allow us to test our construct in a more relevant setting for confronting real-world antibiotic resistance.
Twist: Outreach is an important part of iGEM. What did you do to engage the public with your project?
CAPS Kansas: Over the past year, our iGEM team has designed and presented a number of science events for middle school students in our school district. Each event has attracted between 100-120 students. At these events, participants experience a variety of hands-on activities and demonstrations. Our most recent event in early October was titled “Magical Molecular Adventures”, where we presented activities using Harry Potter as our theme.
Twist: Has synthetic biology changed your view of the world?
CAPS Kansas: Most certainly! Synthetic biology allows us to consider solutions to society’s challenges using mechanisms never before contemplated. With great challenges come great opportunities and synthetic biology provides a much more robust and diverse tool chest to take advantage of these opportunities.
“Synthetic biology allows us to consider solutions to society’s challenges using mechanisms never before contemplated.”
Twist: If you had unlimited access to synthetic DNA, what would you do to change to world?
CAPS Kansas: Given the thrill we have had these past months engaging our project, we would work with younger students in our middle schools to help them develop and execute their own projects with the goal that they too may catch the synthetic biology bug!
Twist: Any final words?
CAPS Kansas: Besides considerable thanks to Twist Bioscience for their sponsorship of our team, we’d also like to thank Desktop Genetics for providing us advice on using their online system for designing our guide RNAs.
Thank you very much to the Blue Valley Schools CAPS iGEM team for taking the time to speak with us. We congratulate you on your accomplishments and wish you success at the 2016 Giant Jamboree competition! Please follow their iGEM team @CAPSiGEM and the CAPS Bioscience program @CAPSBio on Twitter.