Transgenic Drosophila lines for LexA-dependent gene and growth regulation

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 exist...

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Published in:G3 : genes - genomes - genetics 2022-03, Vol.12 (3)
Main Authors: Chang, Kathleen R, Tsao, Deborah D, Bennett, Celine, Wang, Elaine, Floyd, Jax F, Tay, Ashley S Y, Greenwald, Emily, Kim, Ella S, Griffin, Catherine, Morse, Elizabeth, Chisholm, Townley, Rankin, Anne E, Baena-Lopez, Luis Alberto, Lantz, Nicole, Fox, Elizabeth, Kockel, Lutz, Kim, Seung K, Park, Sangbin
Format: Article
Language:English
Subjects:
Animals
Animals, Genetically Modified
Drosophila - genetics
Drosophila - metabolism
Drosophila melanogaster - genetics
Drosophila melanogaster - metabolism
Drosophila Proteins - genetics
Drosophila Proteins - metabolism
Humans
Investigation
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Summary: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.
ISSN:2160-1836
2160-1836
DOI:10.1093/g3journal/jkac018