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Tools for synchronous temporal control of independent transgenes in discrete Drosophila tissues

Evelyne RuchtiiD, Brian D. McCabeiD

Abstract

Exogenous binary gene regulatory expression systems are a linchpin technology critical for many model organism genetic manipulations including of Drosophila melanogaster. Subsequent to the initial establishment of the Gal4/UAS binary expression system, LexA/LexOp and QF/QUAS binary gene expression systems have been adopted to supplement and expand the tissue specific control of genes expression in Drosophila. Here, we have developed a compendium of modular vectors that enable robust, reproducible, and high throughput parallel construction of transgenes that produce either Gal4, LexA or QF2 transcription factors under specific gene enhancers and compatible partner vectors that allow transgenes to be regulated by these factors under UAS, LexOp, or QUAS control. Expanding upon this foundation, we have generated a novel hybrid binary gene regulatory system derived from QF and Gal4 - QFG4, that enables simultaneous coordinate regulation of UAS and QUAS transgenes by Gal80, allowing synchronous temporal control of independent transgene expression in distinct cells or tissues. We envision these new tools will facilitate novel and increasingly sophisticated interrogation of cell and tissue interactions particularly in contexts where temporal control is paramount.

02 /Specification & Classification

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