GAL4 drivers expression in the whole adult fly

Seroude, Laurent

doi:10.1002/gene.10128

Cited by 19 publications

(20 citation statements)

References 20 publications

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“…Although the design of these experiments prevents a rigorous statistical analysis of the changes in the expression of the UAS-lacZ transgene between ages, it is obvious that the expression profile is dependent on the strain, inducer concentration, age and often the gender. Similar to standard GAL4 enhancer-trap strains, a variety of profiles is observed (Seroude, 2002;Seroude et al ., 2002). In addition, within a strain the concentration of the inducer can also influence the profile of lacZ expression.…”

Section: Induction Throughout Life: Expression Levelsmentioning

confidence: 88%

“…In the context of aging studies, it is crucial to ensure that the system does not allow any gene expression in absence of inducer during pre-adult stages. As a stringent test of leakiness (Seroude, 2002), each Gene-Switch driver was combined with a UAS-Grim reporter (encoding a strong pro-apoptotic factor) and embryonic, larval and pupal lethality was scored in absence of inducer and compared to the negative control carrying the reporter alone. Lethality was also measured in presence of 50 μg mL -1 of RU486 to confirm that Grim expression would indeed cause developmental lethality.…”

Section: Pre-adult Expression In Absence Of Inducermentioning

confidence: 99%

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Characterization of the Drosophila Gene‐Switch system in aging studies: a cautionary tale

et al. 2008

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SummaryGenetic studies have shown that in many model organisms, single gene mutations can dramatically influence aging. Systems that allow researchers to control a gene's temporal and spatial expression pattern, known as inducible gene-expression systems, are a valuable asset for the study of the influence of single genes on aging. One inducible gene-expression system reported to allow temporal and tissue-specific control of gene expression in Drosophila is the Gene-Switch system. However, this system has not been extensively characterized in the context of aging research. This report uses six Gene-Switch strains to examine the tissue localization and amount of expression achievable in the major tissue types of the fly. The quantitative analysis of adult flies fed with inducer through life reveals that the levels of expression are influenced by both the inducer concentration and the age of the animal in a strain-specific manner. Furthermore, the relationship between inducer concentration and expression level is unique to each strain and, in some cases, to each gender. The analysis of the spatial expression patterns in several strains revealed expression in more tissue types than previously assumed. Finally, most Gene-Switch strains display expression in the absence of inducer during development and/or during adulthood. These findings have important implications that may reconcile contradictions reported in studies investigating the effects of dFOXO on longevity. This study is an important guide to the design and interpretation of aging studies based on the Gene-Switch system.

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Section: Induction Throughout Life: Expression Levelsmentioning

confidence: 88%

Section: Pre-adult Expression In Absence Of Inducermentioning

confidence: 99%

Characterization of the Drosophila Gene‐Switch system in aging studies: a cautionary tale

et al. 2008

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“…LacZ and GFP enhancer traps are riddling the genome; alternatively, Gal4 traps can be coupled to UAS-reporters of choice to assess gene induction [94]. Below we outline immune-specific reporter strains (listed in Table 2) and how to use them for immune gene quantification.…”

Section: Reporter Linesmentioning

confidence: 99%

Methods to study Drosophila immunity

Neyen

Bretscher

Binggeli

et al. 2014

Methods

132

127

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a b s t r a c tInnate immune mechanisms are well conserved throughout evolution, and many theoretical concepts, molecular pathways and gene networks are applicable to invertebrate model organisms as much as vertebrate ones. Drosophila immunity research benefits from an easily manipulated genome, a fantastic international resource of transgenic tools and over a quarter century of accumulated techniques and approaches to study innate immunity. Here we present a short collection of ways to challenge the fruit fly immune system with various pathogens and parasites, as well as read-outs to assess its functions, including cellular and humoral immune responses. Our review covers techniques for assessing the kinetics and efficiency of immune responses quantitatively and qualitatively, such as survival analysis, bacterial persistence, antimicrobial peptide gene expression, phagocytosis and melanisation assays. Finally, we offer a toolkit of Drosophila strains available to the research community for current and future research.

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“…In the GAL4/UAS system, a tissue specific promoter or enhancer drives expression of the yeast transcription factor GAL4 in one transgenic construct. GAL4 then binds to an upstream activating sequence (UAS) site in the promoter of a second target construct and activates transcription (Brand and Perrimon, 1993;Seroude, 2002;Seroude et al, 2002). Stebbins and coworkers combined these two systems by creating a bridge construct where UAS promoter drives expression of rtTAM2-alt (Stebbins et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Alteration of Drosophila life span using conditional, tissue-specific expression of transgenes triggered by doxycyline or RU486/Mifepristone

Ford

Hoe

Landis

et al. 2007

Experimental Gerontology

117

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The conditional systems Tet-on and Geneswitch were compared and optimized for the tissue-specific expression of transgenes and manipulation of life span in adult Drosophila. Two versions of Tet-on system reverse-tetracycline-Trans-Activator (rtTA) were compared: the original rtTA, and rtTAM2-alt containing mutations designed to optimize regulation and expression. The rtTAM2-alt version gave less leaky expression of target constructs in the absence of doxycycline, however the absolute level of expression that could be achieved was less than that produced by rtTA, in contrast to a previous report. Existing UAS-rtTAM2-alt insertions were re-balanced, and combined with several tissue-general and tissue-specific GAL4 driver lines to yield tissue-specific, doxycycline-inducible transgene expression over three orders of magnitude. The Geneswitch (GS) system also had low background, but the absolute level of expression was low relative to Tet-on. Consequently, actin5C-GS multi-insert chromosomes were generated and higher-level expression was achieved without increased background. Moderate level over-expression of MnSOD has beneficial effects on life span. Here high-level over-expression of MnSOD was found to have toxic effects. In contrast, motor-neuron-specific over-expression of MnSOD had no detectable effect on life span. The results suggest that motor-neuron tissue is not the essential tissue for either MnSOD induced longevity or toxicity in adult males.

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GAL4 drivers expression in the whole adult fly

Cited by 19 publications

References 20 publications

Characterization of the Drosophila Gene‐Switch system in aging studies: a cautionary tale

Characterization of the Drosophila Gene‐Switch system in aging studies: a cautionary tale

Methods to study Drosophila immunity

Alteration of Drosophila life span using conditional, tissue-specific expression of transgenes triggered by doxycyline or RU486/Mifepristone

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