Caffeinated Coli Education Kit We are thrilled to celebrate our talented students, Connie Zhang, Neil Tian, and Riya Reddy, for their outstanding work in developing the Caffeinated Coli Education Kit, which is now being used in select public high schools across Texas! Huge congratulations to them for submitting their manuscript to The Science Teacher —a well-deserved accomplishment! Overview: The Caffeinated Coli Education Kit is a five-week module designed for high school students to learn about synthetic biology, scientific research, and genetic engineering through inquiry-driven learning. The module is divided into: lecture-based classes with student activities and a lab portion where students use a genetically engineered E. coli to measure the caffeine concentration of solutions. Our program is currently being implemented in several Texas high schools, encouraging STEM learning and incorporating novel biology research into high school classes. Learning objectives: Students explore the basics of synthetic biology and research. Students learn fundamental lab skills such as micropipetting, growing bacterial cultures, and conducting a bioassay. Students learn to collect data and analyze data using data analysis software. Breakdown of module: Figure by Neil Tian and Riya Reddy. Background Information: In the wet lab portion of our module, students utilize a strain of Escherichia coli known as ΔguaB E. coli, originally engineered by the UT Austin iGEM 2012 team. This strain of E. coli had their guaB gene knocked out, a gene necessary for de novo guanine synthesis. Without guanine, the bacteria cannot replicate their DNA replication and grow. Therefore, the bacteria were transformed with the pDCAF plasmid, which contains the necessary genes to encode three demethylase enzymes (NdmA, NdmB, and NdmC). These enzymes allow the bacteria to demethylate caffeine (1,3,7-trimethylxanthine) into xanthine. Through the xanthine salvage pathway, the engineered E. coli can convert demethylated xanthine into guanine, restoring their ability to replicate and grow, thus creating a biosensor for caffeine and other methylxanthines. Resources: Download the colorimeter app: https://apps.apple.com/us/app/colorimeter-app/id1542365656 Gutierrez, A, et al.(2019). https://doi.org/10.1128/aem.01965-19 Tian, N, et al. (2025) Caffeinated Coli Educational Kit, submitted.
