ATTRIBUTIONS

We wholeheartedly thank all of the following individuals and institutions for their interest, enthusiasm, and gracious contributions to the development of the CRISPR Capsules project. Their contributions to the project are commendable, and the extent to which their expertise enriched our understanding of synthetic biology is more greatly treasured.

All lab work was completed on the Northwestern University Evanston campus by the members of Northwestern iGEM 2016 unless otherwise noted.

General Support

Our faculty advisers Drs. Mike Jewett, Joshua Leonard, and Keith Tyo talked us through our arduous brainstorming process and guided us as we worked towards our project vision. They also helped the iGEM members in Northwestern’s McCormick School of Engineering and Applied Science obtain grants for living in Evanston over the summer.

Our outstanding graduate student advisors Bradley Biggs, Will Bothfeld, Benjamin Des Soye, Taylor Dolberg, Patrick Donahue, Quentin Dudley, Joseph Muldoon, Kelly Schwarz, Sara Stainbrook, and Peter Su donated time out of their busy lives to discuss our project with us and guide us through our lab experience.

Dr. John Mordacq, lab director and professor of Northwestern University’s Program in Biological Sciences, obligingly accommodated us with a lab space and necessary equipment for our summer experience. He also helped the iGEM members in Northwestern’s Weinberg College of Arts and Sciences obtain grants for traveling and living in Evanston over the summer.

Starting NU iGEM 2016

In February 2016, Northwestern iGEM 2016 was born in a library study room in the presence of chocolate chip cookies and the enthusiastic faces of members of the Northwestern iGEM 2014 team. Thank you for recruiting us and kickstarting an accomplished team dynamic that evolved into staunch friendship.

We would like to thank our phenomenal graduate student TAs for their guidance and dedication to our “bootcamps,” our beginning lab protocol tutorial workshops. Special thanks to Kelly Schwarz, Patrick Donahue, Peter Su, Bradley Biggs, Will Bothfeld, Sara Stainbrook, and Quentin Dudley for walking us through basic lab procedures throughout the spring term before we officially began independent work.

We met with our faculty advisers on a weekly basis during the spring term to zone in on a project direction. Thank you, Drs. Mike Jewett, Joshua Leonard, and Keith Tyo for your mentorship and willingness to discuss project ideas throughout our long brainstorming process.

Lab Support

Technique and Troubleshooting

Thank you to all of the graduate students of the Leonard, Tyo, Jewett, and Bagheri labs who jumped on board to talk to us about our project and help us over the summer!

Kelly Schwarz met with us almost every day to talk to us about our project progress, troubleshooting, and handling the MTAs between our university and Cornell and MIT. She made sure we were caught up on deadlines and gave us our first lesson on how to pour culture plates during the iGEM “bootcamps.” She was unbelievably prompt when we asked her questions through email and kindly provided us with encouraging words whenever we found ourselves in a rut. To the best TA we could have asked for, thank you for your infinite knowledge and patience throughout the summer. We couldn’t have done it without you.

Patrick Donahue walked us through running golden gate assemblies, sending DNA off for sequencing, and helped us troubleshoot our transformations by letting us run streak plate tests with him in Dr. Leonard’s lab. During one of our Bootcamp sessions, he taught us how to run a gel and how to run a restriction digest protocol.

Bradley Biggs taught us how to use Genome Compiler to design our constructs and advised us through the weeks-long process of our first designs. He also helped us troubleshoot our Gibson assembly reactions. During our Bootcamps, he walked us through running our first transformation protocol.

Peter Su helped us make calculations for our first Gibson assembly. He, along with Patrick, taught us how to run gels and restriction digest procedures.

Sara Stainbrook ran our first Bootcamp. She walked us through our first PCR reaction and cleanup, gel imaging, and mixing buffers/reagents.

Will Bothfeld helped us run our first transformation protocol during a Bootcamp session. He also developed a cloning log that helped us determine insert-to-backbone ratios for our Gibson assemblies.

Joseph Muldoon sat in on our weekly meetings and discussed troubleshooting and project progress with us.

Quentin Dudley walked us through our first plasmid miniprep procedure and making glycerol stocks during one of our Bootcamps. During this Bootcamp session he showed us the basics of optical density spectrophotometry.

Benjamin Des Soye graciously ran our western blots with supplies from Dr. Jewett’s lab. He walked us through each step and explained how each part of the procedure worked. He also demonstrated the use of an electroporator when it was needed for transformations.

Taylor Dolberg helped us make stocks of chemically competent cells.

Materials

Thank you to the labs of Drs. Jewett, Leonard, and Tyo for providing us with some reagents to start off our research and to keep us going through late-night experiments, and to Drs. Mordacq and Huaru Yan, who provided us with lab equipment and troubleshooting advice.

Dr. Matthew DeLisa from Cornell University provided us with a hypervesicular strain of E. coli (JC8031) and a His-ClyA-GFP plasmid. The hypervesicular strain is sourced (with permission) from the lab of Dr. Roland Lloubes from the Institute of Microbiology of the Mediterranean.

The lab of Dr. Feng Zhang from the Broad Institute provided us with a plasmid for S. pyogenes Cas9.

Additional Support

Thank you to Dr. Danielle Tullman-Ercek, who advised us on effective periplasm-directing translocation pathways to utilize in our device construction, and to Dr. Michelle Hung, who advised us on working with and purifying vesicles and helped us brainstorm characterization experiments for our project.

Drs. Michael Postelnick, Stephen Persell (MD/MPH), and Marc Sheetz (PharmD) donated their time to answer our questions about the ethical administration of antibiotics despite high public demand for these drugs for our human practices research. Thank you for taking the time to speak with us.

Building with Biology provided us with a means of educating Evanston youth about biotechnology and its applications. They provided us with a kit of activities and exciting discussions we could share with students participating in summer camps at Northwestern. Using our kit from Building with Biology, we were able to partner up with Northwestern University’s Center for Talent Development (CTD) to have an engaging conversation about gene drives. We also gave them a tour of Dr. Jewett’s lab space, where the students made fluorescent bacteria art on culture plates. Northwestern University’s Society of Women Engineers (SWE) hosted its annual Summer Technology and Engineering Program. We talked about biotechnology to the group of girls participating in the event, and facilitated an activity involving isolating DNA from wheat germ using dish soap and rubbing alcohol. The activity was also inspired part of the kit provided by Building with Biology.

Emma Nechamkin from Teach for America allowed us to teach her high school biotechnology class at Perspectives Middle Academy in Auburn Gresham, a neighborhood in the south side of Chicago. Her students had never been exposed to research science, so we were allowed to give them a one-day lecture on CRISPR and antibiotic resistance. Thank you so much for giving us the chance to visit your classroom!

Jessica Yu and Joseph Muldoon helped us troubleshoot our wiki styling problems.

Thank you Drs. Leonard, Tyo and all of our graduate students for helping us with our presentating skills.

Team Roles

Our team emphasizes the integration of skills in all aspect of the CRISPR Capsules project. From wet lab work to speaking at public events, each team member has experience in all fields. Though some of us certainly deem ourselves “specialized” in some tasks, we find that the overall contribution from the high-level development of the project to the technical details of lab troubleshooting constitutes efforts from each member of the team.