e-Article
Transgenic Drosophila lines for LexA-dependent gene and growth regulation.
Document Type
Article
Author
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; Seung K. Kim; Sangbin Park
Source
Subject
*GENETIC regulation
*REGULATION of growth
*DROSOPHILA
*SECONDARY school curriculum
*GENE silencing
*CELL communication
*GENOME editing
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Language
ISSN
2160-1836
Abstract
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 genetictools and teaching methods presented here provide LexA/LexAop resources that complement existing resources to study intercellular communication coordinating metazoan physiology and development. [ABSTRACT FROM AUTHOR]