Expanding the Drosophila toolkit for dual control of gene expression

Zirin, Jonathan; Jusiak, Barbara; Lopes, Raphael; Ewen-Campen, Benjamin; Bosch, Justin A; Risbeck, Alexandria; Forman, Corey; Villalta, Christians; Hu, Yanhui; Perrimon, Norbert

doi:10.7554/elife.94073

Cited by 2 publications

(1 citation statement)

References 44 publications

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“…Recently, a number of new genetic methods have been developed to control cell elimination in a mostly temperature-independent manner. These include GAL80-auxin as well as the DUAL control method (McClure et al, 2022;Zirin et al, 2024;Harris et al, 2020). Using GAL80-auxin, induction of UAS-transgene expression occurs after 18 hours of auxin feeding (McClure et al, 2022), whereas in the DUAL control system LexAop-transgene expression is induced within six hours, however achieving the desired degree of tissue ablation can be limited by the toxicity caused by long heat shocks (Harris et al, 2020).…”

Section: Discussionmentioning

confidence: 99%

Spatiotemporal control of cell ablation using Ronidazole with Nitroreductase inDrosophila

Teeters,

Cucolo,

Kasar

et al. 2024

Preprint

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The ability to induce cell death in a controlled stereotypic manner has led to the discovery of evolutionary conserved molecules and signaling pathways necessary for tissue growth, repair, and regeneration. Here we report the development of a new method to genetically induce cell death in a controlled stereotypic manner in Drosophila. This method has advantages over other current methods and relies on expression of the E. coli enzyme Nitroreductase (NTR) with exogenous application of the nitroimidazole prodrug, Ronidazole. NTR expression is controlled spatially using the GAL4/UAS system while temporal control of cell death is achieved through timed feeding of Ronidazole supplied in the diet. In cells expressing NTR, Ronidazole is converted to a toxic substance inducing DNA damage and cell death. Caspase cell death is achieved in a range of NTR-expressing cell types with Ronidazole feeding, including epithelial, neurons, and glia. Removing Ronidazole from the diet restores cell death to normal unperturbed levels. Unlike other genetic ablation methods, temporal control is achieved through feeding not temperature, circumventing developmental complications associated with temperature changes. Ronidazole-NTR also requires only two transgenes, a GAL4 driver and UAS-NTR, which is generated as a GFP-NTR fusion allowing for easy setup of large-scale screening of UAS-RNAi lines. Altogether, Ronidazole-NTR provides a new streamlined method for inducing cell death in Drosophila with temperature-independent ON/OFF control.

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Section: Discussionmentioning

confidence: 99%

Spatiotemporal control of cell ablation using Ronidazole with Nitroreductase inDrosophila

Teeters,

Cucolo,

Kasar

et al. 2024

Preprint

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EyaHOST, a modular genetic system for investigation of intercellular and tumor-host interactionsin Drosophila melanogaster

Teles-Reis,

Jain,

Liu

et al. 2024

Preprint

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Cell biology and genetic analysis of intracellular, intercellular and inter-organ interaction studies in animal models are key for understanding development, physiology, and disease. The MARCM technique can emulate tumor development by simultaneous clonal tumor suppressor loss-of-function generation coupled with GAL4-UAS-driven oncogene and marker expression, but the utility is limited for studying tumor-host interactions due to genetic constraints. To overcome this, we introduce EyaHOST, a novel system that replaces MARCM with the QF2-QUAS binary gene expression system under theeyapromoter control, unleashing the fly community genome-wide GAL4-UAS driven tools to manipulate any host cells or tissue at scale. EyaHOST generates epithelial clones in the eye epithelium similar to MARCM. EyaHOST-driven RasV12oncogene overexpression coupled with scribble tumor suppressor knockdown recapitulates key cancer features, including systemic catabolic switching and organ wasting. We demonstrate effective tissue-specific manipulation of host compartments such as neighbouring epithelial cells, immune cells, fat body, and muscle using fly avatars with tissue-specific GAL4 drivers. Organ-specific inhibition of autophagy or stimulation of growth-signaling through PTEN knockdown in fat body or muscle prevents cachexia-like wasting. Additionally, we show thatRasV12, scribRNAitumors induce caspase-driven apoptosis in the epithelial microenvironment. Inhibition of apoptosis by p35 expression in the microenvironment promotes tumor growth. EyaHOST offers a versatile modular platform for dissecting tumor-host interactions and other mechanisms involving intercellular and inter-organ communication inDrosophila.Highlights*eyes absent, eye disc-specific enhancer drives clonal KD recombinase flip-out activated QF2 expression in the larval eye epithelium for simultaneous QUAS-driven gain and loss-of-function analysis of gene function.*Clones are visualized by QUAS-tagBFP or QUAS-eGFP facilitating analysis of existing fluorescent reporters.*The GAL4-UAS system and existing genome-wide genetic tools are released to independently manipulate any cell population in the animal for cell biology, intercellular or inter-organ analysis for developmental, physiological, or disease model analysis.*Fly avatars for tumor-host interaction studies with multiple organs allow live monitoring and manipulation of tumors and organs in translucent larva.

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Expanding the Drosophila toolkit for dual control of gene expression

Cited by 2 publications

References 44 publications

Spatiotemporal control of cell ablation using Ronidazole with Nitroreductase inDrosophila

Spatiotemporal control of cell ablation using Ronidazole with Nitroreductase inDrosophila

EyaHOST, a modular genetic system for investigation of intercellular and tumor-host interactionsin Drosophila melanogaster

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