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CRISPR/Cas9 in Yeast as a Tool for Teaching and Reinforcing Molecular Biology Concepts to Undergraduates

Undergraduate students in an upper‐level molecular biology laboratory class used their training in biochemistry, cell biology and molecular biology to design and perform CRISPR/Cas9 genome editing on yeast. After an introduction to DNA repair and CRISPR/Cas9 in the lecture, the students took on the task to design an experiment using Cas9 to generate auxotrophic yeast mutants. They used readily available bioinformatics databases to retrieve yeast genome sequence and design CRISPR‐associated sgRNA to target the gene of interest. They cloned genes for the sgRNAs into a commercially available CRISPR/Cas9 vector, and then transformed it with a template for homology‐directed repair into yeast. Next, the students phenotyped and genotyped the potential auxotrophic mutants generated by CRISPR/Cas9. As the penultimate laboratory course in the program, this lab introduced new skills, provided opportunity to apply concepts discussed in lecture, and reinforced skills and knowledge from previous courses. A survey, exam and formal lab report assessed positive outcomes in learning and affect. The reagents in this laboratory course design are readily available and financially feasible for undergraduate laboratory courses, and the protocols/emphasis are adaptable to diverse program goals. Overall, incorporating the tractable, yet modern gene editing technology into undergraduate laboratory courses is both realistic and favorable to learning.