Abstract
Training the future synthetic biology workforce requires the opportunity for students to be exposed to biotechnology concepts and activities in secondary education. Detecting Wolbachia bacteria in arthropods using polymerase chain reaction (PCR) has become a common way for secondary students to investigate and apply recombinant DNA technology in the science classroom. Despite this important activity, cutting-edge biotechnologies such as clustered regularly interspaced short palindromic repeat (CRISPR)-based diagnostics have yet to be widely implemented in the classroom. To address this gap, we present a freeze-dried CRISPR-Cas12 sensing reaction to complement traditional recombinant DNA technology education and teach synthetic biology concepts. The reactions accurately detect Wolbachia from arthropod-derived PCR samples in under 2 h and can be stored at room temperature for over a month without appreciable degradation. The reactions are easy-to-use and cost less than $40 to implement for a classroom of 22 students including the cost of reusable equipment. We see these freeze-dried CRISPR-Cas12 reactions as an accessible way to incorporate synthetic biology education into the existing biology curriculum, which will expand biology educational opportunities in science, technology, engineering, and mathematics.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 835-842 |
| Number of pages | 8 |
| Journal | ACS synthetic biology |
| Volume | 11 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 18 2022 |
Funding
We would like to acknowledge miniPCR bio for their contribution of materials and reagents. We would also like to thank Sarah Bordenstein and the Wolbachia Project for helpful conversation and for N. vitripennis specimens. We would also like to acknowledge the Centennial High School DNA Club for providing collaborative biology research opportunities for high school students. We would like to acknowledge members of the Jewett Lab for helpful discussions. M.C.J. acknowledges support from the Department of Energy grants DE-SC0018249, DE-NA0003525, and 8J-30009-0029A, the DOE Joint Genome Institute ETOP program, the Office of Energy Efficiency and Renewable Energy grant DE-EE0008343, the David and Lucile Packard Foundation, the Camille Dreyfus Teacher-Scholar Program, the Defense Threat Reduction Agency grants HDTRA1-15-10052/P00001, HDTRA-12-01-0004, and HDTRA12110038, the Army Research Office grants W911NF-20-1-0195, W911NF-18-1-0200, and W911NF-16-1-0372, the National Science Foundation grants 1936789 and 1844336, the Asian Office for Air Force Research Grant FA2386-21-1-4078, and the Army Contracting Command Contract W52P1J-21-9-3023. The work conducted by the U.S. Department of Energy Joint Genome Institute, a DOE Office of Science User Facility, was supported by the Office of Science of the U.S. Department of Energy under contract no. DE-AC02-05CH11231. G.A.R was supported by the National Science Foundation Graduate Research Fellowship Program under grant no. DGE-1842165.
Keywords
- CRISPR
- Cas12
- Wolbachia
- nucleic acid detection
- secondary science education
ASJC Scopus subject areas
- Biomedical Engineering
- Biochemistry, Genetics and Molecular Biology (miscellaneous)