Chang, K.R., Tsao, D.D., Bennett, C., Wang, E., Floyd, J.F., Tay, A.S.Y., Greenwald, E., Kim, E.S., Griffin, C., Morse, E., Chisholm, T., Rankin, A.E., Baena-Lopez, L.A., Lantz, N., Fox, E., Kockel, L., Kim, S.K., Park, S. (2022). Transgenic Drosophila lines for LexA-dependent gene and growth regulation.  G3 (Bethesda) 12(3): jkac018.

FBrf0252870

Research paper

Conditional expression of short hairpin RNAs with binary genetic systems is an indispensable tool for studying gene function. Addressing mechanisms underlying cell-cell communication in vivo benefits from simultaneous use of 2 independent gene expression systems. To complement the abundance of existing Gal4/UAS-based resources in Drosophila, we and others have developed LexA/LexAop-based genetic tools. Here, we describe experimental and pedagogical advances that promote the efficient conversion of Drosophila Gal4 lines to LexA lines, and the generation of LexAop-short hairpin RNA lines to suppress gene function. We developed a CRISPR/Cas9-based knock-in system to replace Gal4 coding sequences with LexA, and a LexAop-based short hairpin RNA expression vector to achieve short hairpin RNA-mediated gene silencing. We demonstrate the use of these approaches to achieve targeted genetic loss-of-function in multiple tissues. We also detail our development of secondary school curricula that enable students to create transgenic flies, thereby magnifying the production of well-characterized LexA/LexAop lines for the scientific community. The genetic tools and teaching methods presented here provide LexA/LexAop resources that complement existing resources to study intercellular communication coordinating metazoan physiology and development.

PMC8895989 (PMC) (EuropePMC)