Gmr gal4

Manufactured by Bloomington Drosophila Stock Center

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GMR-Gal4 is a genetically modified Drosophila melanogaster line that expresses the Gal4 transcription factor under the control of the Glass Multimer Reporter (GMR) promoter. This promoter drives expression in the developing and mature compound eye of the fly. The GMR-Gal4 line can be used to direct the expression of transgenes to the Drosophila eye.

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GMR-GAL4) (BDSC#1104) (2 citations)

79 protocols using gmr gal4

Drosophila Genetic Manipulation Techniques

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Fly stocks and crosses were cultured on a standard medium composed of cornmeal, molasses, yeast, and agar, and treated with propionic acid and methylparaben to inhibit fungal growth. The stocks were raised at 23° ± 2°C, while the crosses and experiments were performed at 25° and 29°C, respectively. The CG4495 RNA interference lines, w 1118 ; P{GD4927}v49349 and w 1118 ; P{GD4927}v49350, hereafter referred to as UAS-MICU1-RNAi ( 1) and UAS-MICU1-RNAi (2), respectively, were obtained from the Vienna Drosophila Resource Center (Vienna, Austria). The UAS-Buffy (Quinn et al., 2003) (link) was kindly provided by Dr. L. Quinn (University of Melbourne, Melbourne, Australia) and Ddc-Gal4 flies (Li et al., 2000) (link) by Dr. J. Hirsch (University of Virginia, USA). GMR-Gal4 (Freeman, 1996) (link) and UAS-lacZ flies were obtained from the Bloomington Drosophila Stock Center (Indiana University, USA).
The UAS-Buffy/CyO; Ddc-Gal4 and UAS-Buffy/CyO; GMR-Gal4 derivative lines were generated by using the standard homologous recombination methods, and were used for overexpression of Buffy in neurons using the Ddc-Gal4 transgene or in the developing eye using the Glass Multiple Reporter (GMR) response elements following a standard protocol described previously (M'Angale and Staveley, 2016b, c) .

M'Angale P.G, & Staveley B.E. (2017). Inhibition of mitochondrial calcium uptake 1 in Drosophila neurons. Genetics and molecular research : GMR, 16(1).

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Drosophila Genetic Manipulation Protocol

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Flies were maintained at 25 °C on standard fly food. The fly strains GMR-Gal4 (#1104), Tub-Gal80^ts (Bloomington 7019), Tub-Gal4 (Bloomington 5138), and c739-Gal4,UAS-mCD8::GFP (#64305) were obtained from the Bloomington Stock Center, while 40D-UAS (#60101) for the control experiment was purchased from VDRC (Vienna, Austria).

Itai T., Sugie A., Nitta Y., Maki R., Suzuki T., Shinkai Y., Watanabe Y., Nakano Y., Ichikawa K., Okamoto N., Utsuno Y., Koshimizu E., Fujita A., Hamanaka K., Uchiyama Y., Tsuchida N., Miyake N., Misawa K., Mizuguchi T., Miyatake S, & Matsumoto N. (2023). A novel NONO variant that causes developmental delay and cardiac phenotypes. Scientific Reports, 13, 975.

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Ubiquitin Proteomics in Drosophila

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UAS-BirA and UAS-(^BioUb)₆-BirA [39 (link)] and their recombination with GMR-GAL4 flies for the study of ubiquitin proteomics has been previously described [43 (link)]. FLAG-tagged Parkin wild-type (Parkin^WT) and Parkin^LD flies were generated at Bestgene using the pUASattb constructs described above. Both UAS-Parkin^WTand UAS-Parkin^LD lines were independently crossed with GMR-GAL4,UAS-(^BioUb)₆-BirA to finally generate: GMR-GAL4,UAS-(^BioUb)₆-BirA/CyO;UAS-Parkin^WT and GMR-GAL4,UAS-(^BioUb)₆-BirA/CyO;UAS-Parkin^LD. GMR-GAL4/CyO;UAS-BirA/TM6 and GMR-GAL4,UAS-(^BioUb)₆-BirA/CyO flies were additionally used as controls. UAS-GFP, elav-GAL4, GMR-GAL4, Tub-GAL4, Da-GAL4, Ple-GAL4 flies were obtained from Bloomington Drosophila Stock Center. UAS-GFP^CL1 flies were obtained from [48 (link)] and park²⁵/TM6b GFP-w⁺ and UAS-park were obtained from [16 (link)]. Flies were grown in 12 h light-dark cycles at 25 °C and were fed with wheat flour and yeast food (1% agar, 5.5% dextrose, 3.5% wheat flour, 5% yeast, 0.25% Nipagen, 0.4% Propionic acid and 0.02% Benzalkonium Chloride in distilled H₂O).

Martinez A., Lectez B., Ramirez J., Popp O., Sutherland J.D., Urbé S., Dittmar G., Clague M.J, & Mayor U. (2017). Quantitative proteomic analysis of Parkin substrates in Drosophila neurons. Molecular Neurodegeneration, 12, 29.

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Genetic Manipulation of Drosophila Models

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The w¹¹¹⁸ strain was used as the wild-type control. The UAS-PAR-1-WT, UAS-PAR-1-T408 A, and UAS-htauM were described previously (Lee et al., 2012 (link)). The Mhc-GAL4 fly line was provided by Dr. T Littleton. The FAF^EP381, UAS-Pod-1-WT, UAS-Pod-1-RNAi, UAS-Slimb-RNAi, and GMR-GAL4 lines were obtained from Bloomington Drosophila Stock Center. The flies were maintained at 25 °C on standard Drosophila food.

Kang H.Y., Kim H.J., Kim K., Oh S.I., Yoon S., Kim J., Park S., Cheon Y., Her S., Lee M., Lu B, & Lee S. (2020). Actin-microtubule crosslinker Pod-1 tunes PAR-1 signaling to control synaptic development and tau-mediated synaptic toxicity. Neurobiology of aging, 90, 93-98.

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Fly Genetics and Molecular Techniques

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Drosophila melanogaster stocks and crosses were maintained in humidified incubators at 25°C with 12 hr light-dark cycles. The alleles Nab2^ex3 (null), Nab2^pex41 (precise excision 41; control), and UAS-Flag-Nab2 have been described previously (Pak et al., 2011 (link); Kelly et al., 2014 (link)). Lines from Bloomington Drosophila Stock Center (BDSC): GMR-Gal4 (#1350), elav^C155-Gal4 (#458), msl-2²²⁷ (#5871), msl-2^kmA (#25158), mle⁹ (#5873), roX1^ex6 (#43647), UAS-Mettl3, UAS-Mettl3-RNAi (#80450), fl(2)d² (#36302), vir^2F (#77886). The Mettl3^null, UAS-Mettl3, and Ythdc^ΔN alleles were all kind gifts of J-Y. Roignant. The Nab2^ex3,Mettl3^null and Nab2^ex3, Ythdc^ΔN chromosomes were generated by meiotic recombination and confirmed by genomic PCR. A total of 200 recombinant lines were screened to identify Nab2,Mettl3 double mutants.

Jalloh B., Lancaster C.L., Rounds J.C., Brown B.E., Leung S.W., Banerjee A., Morton D.J., Bienkowski R.S., Fasken M.B., Kremsky I.J., Tegowski M., Meyer K., Corbett A, & Moberg K. (2023). The Drosophila Nab2 RNA binding protein inhibits m6A methylation and male-specific splicing of Sex lethal transcript in female neuronal tissue. eLife, 12, e64904.

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Drosophila Genetic Manipulation Protocol

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Drosophila stocks were maintained at 25°C. The GAL4-UAS system (Brand and Perrimon, 1993) was used for gene overexpression or knockdown. Genetic crosses were made at 25°C unless stated otherwise. w¹¹¹⁸ was used as wild-type. GAL4 and UAS lines were: GMR-GAL4 (Bloomington Drosophila Stock Center), UAS-sti RNAi^HMJ03025 (Bloomington Stock Center, BL64012, sti RNAi¹ in short), UAS-sti RNAi^NIG10522R-1 (National Institute of Genetics, NIG 10522R-1, sti RNAi²), UAS-sti RNAi^NIG10522R-2 (NIG 10522R-2, sti RNAi³), UAS-wts RNAi (Vienna Drosophila Research Center, v9928), and UAS-wts (a gift from G. Halder). sti¹, sti³, and sti^GS9053 mutants were obtained from the Bloomington stock center (BL4366, BL4871) and the Kyoto stock Center (DGRC201488), respectively. Eye sizes were measured using the Image J software. Dot plot graphs were generated using the R language in the RStudio IDE with the ggplot2 package.

Tran T.H., Yang D.W., Kim M., Lee D.H., Gai M., Di Cunto F., Choi K.W, & Lim D.S. (2019). Citron kinase interacts with LATS2 and inhibits its activity by occluding its hydrophobic phosphorylation motif. Journal of Molecular Cell Biology, 11(11), 1006-1017.

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Fly Husbandry and Transgenic Lines

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Fly stocks were kept at 25°C on a 12-h light and 12-h dark cycle and fed a standard sugar/yeast/agar diet (Bass et al, 2007 (link)). The light intensity in the fly chambers was around 1,000 lux. The flies were reared at controlled larval densities, and once-mated female flies were used for all experiments unless otherwise stated. The flies were snap-frozen with liquid nitrogen. Dissections were carried out in PBS and tissues either directly analysed or frozen on dry ice.
Transgenic flies were backcrossed into the outbred white Dahomey (wDah) or red Dahomey (rDah) wild-type strain (Grönke et al, 2010 (link)) with the endosymbiont Wolbachia for at least six generations, if necessary. The following transgenic fly lines were used in this study: GMR-Gal4 (Bloomington), UAS-miR-210 (Bejarano et al, 2012 (link)), Sima KO/Tm3Sb (Bloomington #14640), Dgk RNAi (Bloomington #41944), GMR-p35 (Bloomington #5774), Df(1)BSC352 (Bloomington #24376), Apc RNAi (VDRC v51468), Vha55 RNAi (VDRC v46553), Fasn1 RNAi (VDRC v29349) (Dietzl et al, 2007 (link)), UAS-Fasn1 (Garrido et al, 2015 (link)), and UAS-bmm-eGFP (Grönke et al, 2005 (link)).

Weigelt C.M., Hahn O., Arlt K., Gruhn M., Jahn A.J., Eßer J., Werner J.A., Klein C., Büschges A., Grönke S, & Partridge L. (2019). Loss of miR-210 leads to progressive retinal degeneration in Drosophila melanogaster. Life Science Alliance, 2(1), e201800149.

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Transgenic Fly Stock Backcrossing Protocol

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All transgenic fly stocks used, with the exception of w(IR) and GMR-Gal4, were backcrossed into our in-house wild type strain, the Canton-S derivative w¹¹¹⁸ (isoCJ1) [107 (link)], for at least five generations to homogenize genetic background. The GFP, OK107-, ELAV-, and Repo-Gal4 lines [108 (link)], as well as the hTDP-43 [68 (link)] and gypsy(IR) [57 (link)] lines, are as reported previously. The GMR-Gal4, Gal80^ts, w(IR), GFP(IR), and tdTomato lines were acquired from the Bloomington Drosophila Stock Center (stock numbers: 43675, 7019, 25785, 9331, and 32221; respectively), and the loki(IR) line was acquired from the Vienna Drosophila Resource Center [109 (link)] (stock number: v44980). Flies were cultured on standard fly food at room temperature unless otherwise noted.

Krug L., Chatterjee N., Borges-Monroy R., Hearn S., Liao W.W., Morrill K., Prazak L., Rozhkov N., Theodorou D., Hammell M, & Dubnau J. (2017). Retrotransposon activation contributes to neurodegeneration in a Drosophila TDP-43 model of ALS. PLoS Genetics, 13(3), e1006635.

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Drosophila Tau Transgenic Lines

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GMR-GAL4, elav-GAL4, actin-GAL4, tubulin-Gal4, tau^HM05101, tubP-GAL80^ts, UAS-Dcr-2, tau^Df(3R)MR22, Df(3R)BSC499, and UAS-hTau23 (UAS-MAPT.A59A) were obtained from the Bloomington Stock Center. UAS-dTau::GFP and tau^EP3597 were a gift of D. St. Johnston (University of Cambridge, Great Britain). Appl-Gal4 was obtained from Laura Torroja (Universidad Autonoma de Madrid, Spain). tau^GD25023 was received from the Vienna Drosophila RNAi Center (Vienna, Austria). olk¹ mutants are described in (Bettencourt da Cruz et al., 2005 (link)). Flies were raised under standard conditions at 25°C unless specifically described. To create the MAP60 and MAP205-expressing flies, we used LD02709 for MAP60 and LD12965, which were cloned into pUAST. Transgenic flies were generated by P-element transformation using Best Gene Inc. (Chino Hills, CA).

Bolkan B.J, & Kretzschmar D. (2014). Loss of Tau results in defects in photoreceptor development and progressive neuronal degeneration in Drosophila. Developmental neurobiology, 74(12), 1210-1225.

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Drosophila Pupal Eye Developmental Protocol

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All experiments were carried out at 25⁰ C, except heat shocks, which were carried out at 37⁰ C in a water bath. For experiments with staged pupae, white prepupae were marked (counted as 0hr APF) and aged for the appropriate number of hours. Due to the length of the white prepupal stage (Bainbridge and Bownes, 1981 (link)), pupal ages listed may be +20 minutes. Stocks used are as follows (Bloomington Stock numbers are listed where applicable): w¹¹¹⁸, GMR-Gal4 [w^*;P{GAL4-ninaE.GMR}12] #1104, UAS-E2f^FUCCI/UAS-CycB^FUCCI [w¹¹¹⁸;P{UASp-GFP.E2f1.1-230}64, P{UASp-mRFP1.NLS.CycB.1-266}5] #55111, GMR-p21 [y¹ w¹¹¹⁸; P{GMR-p21.Ex}3] #8414, UAS-stg [w¹¹¹⁸; P{UAS-stg.N}4] #4778, CycB-GFP [y¹ w^*; P{PTT-GC}CycB^CC01846] #51568, hs-stg, (Edgar and O’Farrell, 1990 (link)), UAS-p35 [w^*; P{UAS-p35.H}BH2] #5073, hs-Flp, FRT19A Ubi-mRFP.nls [P{Ubi-mRFP.nls}1, w^*, P{hsFLP}12 P{neoFRT}19A] #31418, FRT19A His2Av-GFP [y¹ w^* P{His2Av^{T:Avic\GFP-S65T}}1 P{hsFLP}12 P{neoFRT}19A/FM7a] #32045, UAS-p21 (I. Hariharan). Genotypes listed in figures as GMR-p21, GMR>p21, or GMR>stg (except Fig. 1K) also contain UAS-FUCCI. Transgenic genotypes described in all experiments are heterozygous, with the exception of GMR>FUCCI alone (as in Fig. 1E) and CycB-GFP alone (as in Fig. S1A).

Meserve J.H, & Duronio R.J. (2017). A population of G2-arrested cells are selected as sensory organ precursors for the interommatidial bristles of the Drosophila eye. Developmental biology, 430(2), 374-384.

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