CRISPR/Cas9 gene editing inDrosophilavia visual selection in a summer classroom

Abstract

AbstractCRISPR/Cas9 methods are a powerfulin vivoapproach to edit the genome ofDrosophila melanogaster. To convert existingDrosophila GAL4lines toLexAdriver lines in a secondary school classroom setting, we applied the CRISPR-based genetic approach to a collection ofGal4‘driver’ lines. The integration of theyellow+coat color marker into homology-assisted CRISPR knock-in (HACK) enabled visual selection ofGal4-to-LexAconversions using brightfield stereo-microscopy available in a broader set of standard classrooms. Here, we report the successful conversion of elevenGal4lines with expression in neuropeptide-expressing cells into corresponding, novelLexAdrivers. The conversion was confirmed byLexA-andGal4-specific GFP reporter gene expression. This curriculum was successfully implemented in a summer course running 16 hours/week for seven weeks. The modularity, flexibility, and compactness of this course should enable development of similar classes in secondary schools and undergraduate curricula, to provide opportunities for experience-based science instruction, and university-secondary school collaborations that simultaneously fulfill research needs in the community of science.