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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. © The Author(s) 2022. Published by Oxford University Press on behalf of Genetics Society of America.


Kathleen R Chang, Deborah D Tsao, Celine Bennett, Elaine Wang, Jax F Floyd, Ashley S Y Tay, Emily Greenwald, Ella S Kim, Catherine Griffin, Elizabeth Morse, Townley Chisholm, Anne E Rankin, Luis Alberto Baena-Lopez, Nicole Lantz, Elizabeth Fox, Lutz Kockel, Seung K Kim, Sangbin Park. Transgenic Drosophila lines for LexA-dependent gene and growth regulation. G3 (Bethesda, Md.). 2022 Mar 04;12(3)

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PMID: 35100369

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