CRISPR/Cas9 gene editing in Drosophila via visual selection in a summer classroom

Kockel, L.; Zhang, V.; Wang, J.; Gulick, C.; Laws, M. E.; Rajan, A.; Lantz, N.; Asgarova, A.; Dai, L.; Garcia, K.; Kim, C.; Li, M.; Ordonez-Acosta, P.; Peng, D.; Shull, H.; Tse, L.; Wang, Y.; Yu, W.; Zhou, Z.; Rankin, A.; Park, S.; Kim, S. K.

doi:10.1101/2024.03.28.587232

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bioRxiv · 10.1101/2024.03.28.587232

CRISPR/Cas9 gene editing in Drosophila via visual selection in a summer classroom

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Abstract

CRISPR/Cas9 methods are a powerful in vivo approach to edit the genome of Drosophila melanogaster. To convert existing Drosophila GAL4 lines to LexA driver lines in a secondary school classroom setting, we applied the CRISPR-based genetic approach to a collection of Gal4 driver lines. The integration of the yellow+ coat color marker into homology-assisted CRISPR knock-in (HACK) enabled visual selection of Gal4-to-LexA conversions using brightfield stereo-microscopy available in a broader set of standard classrooms. Here, we report the successful conversion of eleven Gal4 lines with expression in neuropeptide-expressing cells into corresponding, novel LexA drivers. The conversion was confirmed by LexA- and Gal4-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.

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Kockel, L., Zhang, V., Wang, J., Gulick, C., Laws, M. E., Rajan, A., Lantz, N., Asgarova, A., Dai, L., Garcia, K., Kim, C., Li, M., Ordonez-Acosta, P., Peng, D., Shull, H., Tse, L., Wang, Y., Yu, W., Zhou, Z., Rankin, A., Park, S., Kim, S. K.. 2024-03-30. CRISPR/Cas9 gene editing in Drosophila via visual selection in a summer classroom. https://doi.org/10.1101/2024.03.28.587232

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