Team:USAFA/Attributions
Attributions
With 19 biology, biochemistry, and civil engineering majors making up our fearless team of PFAS warriors, the USAFA iGEM team was fully equipped to take on the task of PFAS degradation and detection. Listed below on this page are the specific attributions from our team members as well as collaborations. We would like to thank our advisors, Dr. Jordan Steel, Major Erin Almand, Captain John Sitko, and Dr. Anthony Arment for their mentorship and guidance throughout the project, without them, this project would not have been possible. We would also like to thank all those who collaborated on this project, Dr. Vanessa Varaljay, Dr. Chia Hung, Dr. Rajiv Berry, and Dr. Nancy Kelley-Loughnane from the Air Force Research and Laboratory, Environmental Engineers and PFAS experts Pierre Kwan and Elizabeth Mende, and our fellow Colorado iGEM teams.
Individual Soil Isolates
Jackson Harris = Research head. Growth of microbe library in PFAS minimal media and growth characterization. Identification of dehalogenases from Delftia acidovorans. Biobrick design and cloning. Dehalogenase purification and characterization when exposed to fluorinated compounds. Safety form co-author. Modeling of Dehalogenases and binding PFAS.
Chris Jeon = Measured F ion release using a hemicyanine dye and EDTA complex. Completed the safety form and safety wiki. Collected team pictures and bios for the wiki page.
Meaghan Raab = Extracted RNA and transcriptomics of exposed vs control.
Dehalogenase
Peter Lochmaier = Tested for viability with PFAS and F ion release. Lead for Bioinformatics Research.
Twee Pham = Worked to transform and degrade PFAS using Dehalogenases. Lead for the creative vision, designing team shirts and lab coats, creating several PFAS cartoons, wrote an original PFAS inspired song, and starred in the music video featuring this hit.
Brian Swicegood = Clone the fragment into plasmid vector (Dehalogenase), HP
Megan Doherty = Conducted cell lysis experiment to determine the presence of dehalogenase in bacterial cells by measuring fluoride ion levels over a 24-hour period to determine if PFAS was degraded. Spearheaded many of our collaboration efforts, organizing an interview with civil engineers to gain perspective on current PFAS detection and removal methods, and a virtual meeting with iGEM teams from Colorado School of Mines and University of Colorado Boulder to discuss current project process and COVID-19 impact.
Abbie Loesch = Conducted cell lysis experiments to determine the presence of dehalogenase in bacterial cells by measuring fluoride ion levels over a 24-hour period to determine if PFAS was degraded. HP
Annelise Holland = Conducted cell lysis experiment to determine the presence of dehalogenase in bacterial cells by measuring fluoride ion levels over a 24-hour period to determine if PFAS was degraded. Organizing our experiment in the Cool Science Fair and team gear.
Fungal Isolates
Eamon McHugh = Aimed to discover a fungal exoenzyme with the capability of PFAS degradation. Attempted to culture a variety of fungi from Winogradsky columns and soil samples on minimal PFOA media and has grown three promising fungi.
Alissa Till = Aimed to discover a fungal exoenzyme with the capability of PFAS degradation. Cultured a variety of fungal soil samples in large amounts, three promising fungi were found. Collaborated on the Cool Science Fair experiment for Human Practices.
Detection
Maddie Reicher = Pushed forward with positive results from the 2019 USAFA iGEM team, working to further optimize the prmA promoter, known to upregulate in the presence of PFAS. Created DROP logo and several other graphic designs for the 2020 team.
Olivia Orahood = Worked to find a constitutive promoter within Rhodococcus to use as a positive control for RFP expression and move towards a detection method for PFAS. Collaborated on the Cool Science Fair experiment for Human Practices.
Conley Walters = Experimented with a dehalogenase promoter as a form of detection for PFAS. Collaborated on the Cool Science Fair experiment for Human Practices.
Advisors
Dr. Jordan Steel = Head Team Advisor. Dehalogenase Team Mentor.
Major Erin Almand = Winogradsky Community, Fungus Isolate Team Mentor.
Captain John Sitko = Detection Team Mentor. Edited film for team music video and trailer.
Dr. Anthony Arment = Individual Soil Isolates Team Mentor
Special Thanks
General Support = Patti Kryzanowski, USAFA Biology Lab Manager. Morgan Vance USAFA Biology Equipment Specialist.
Project Support and Advice: USAFA Faculty: Dr. Jordan Steel, Major Erin Almand, Captain John Sitko, Dr. Anthony Arment and AFRL researchers: Dr. Vanessa Varaljay, Dr. Chia Hung, Dr. Rajiv Berry, and Dr. Nancy Kelley-Loughnane
Fundraising/Funding Help: Dr. Don Veverka, Life Science Research Center USAFA and Dr. Nereyda Sevilla, Defense Health Agency Clinical Investigations Program
Lab Support: Patti Kryzanowski, USAFA Biology Lab Manager.
Difficult Technique Support: Dr. Rajiv Chia Hung, Modeling Techniques.
Wiki Support: C2C Eamon McHugh.
Presentation Coaching: USAFA Biology Department Faculty.
Human Practices Support: Major Erin Almand.
Working every other day in Bio 365, the Air Force Academy’s synthetic biology course, the USAFA iGEM 2020 team began our project in January of this year. Our syllabus and course materials are attached below. The PFAS detection and degradation project began in 2019, with pFAST, and this year’s team built off pFAST results with DROP, brainstorming more innovative ways to detect and degrade PFAS, beginning back in November 2019. Work in the lab for DROP began in February of this year, cut short by COVID and continued back up again in August.