Ok107 gal4

Manufactured by Bloomington Drosophila Stock Center

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OK107-GAL4 is a Drosophila melanogaster (fruit fly) genetic tool used to drive gene expression in a specific subset of neurons. It is a GAL4 driver line that expresses the yeast transcriptional activator GAL4 under the control of the OK107 enhancer, targeting expression to the mushroom body, a brain region involved in learning and memory.

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13 protocols using ok107 gal4

Rearing Drosophila Flies for Genetic Studies

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Flies were reared on cornmeal/molasses/agar medium under standard culture conditions (29°C, 25°C or 18°C depending on the presence of tubP-Gal80^ts, 12 h:12 h light/dark cycle). CO₂ was used as an anesthetic. slgA^NP4104, UAS-mCD8-gfp, UAS-mito-tomato, UAS-RFP, CkIIα^JF01436, CkIIα ^GL0003, slgA^GL01514, TubP-Gal80^ts, TubP-Gal4, OK107-Gal4 and 201y-Gal4 were obtained from the Bloomington Drosophila Stock Center, Bloomington, IN, USA. P{cry-Gal4.E39} and P{pdf-Gal4.P2.4} were a gift of Dr Bassem Hassan (ICM, Paris, France). All fly stocks were isogenized by mating females to Canton-S males for ten generations to exclude effects due to differences in genetic background. The RNAi lines used were predicted to have no off-target effects (Ni et al., 2009 (link)).

Zwarts L., Vulsteke V., Buhl E., Hodge J.J, & Callaerts P. (2017). SlgA, encoded by the homolog of the human schizophrenia-associated gene PRODH, acts in clock neurons to regulate Drosophila aggression. Disease Models & Mechanisms, 10(6), 705-716.

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Drosophila Tau and Amyloid-Beta Models

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Flies were raised at a standard density with a 12 h:12 h light dark (LD) cycle with lights on at ZT 0 (Zeitgeber time) on standard Drosophila medium (0.7% agar, 1.0% soya flour, 8.0% polenta/maize, 1.8% yeast, 8.0% malt extract, 4.0% molasses, 0.8% propionic acid, and 2.3% nipagen) at 25°C. The following flies used in this study were previously described or obtained from the Bloomington and Vienna fly stock centers: wild type control was Canton S w- (CSw-) (gift from Dr. Scott Waddell, University of Oxford). Experimental genotypes were elav-Gal4 (Bloomington stock center line number BL8760), OK107-Gal4 (BL854), GMR-Gal4 (BL9146), uas-human MAPT (TAU 0N4R) wild type (gift from Dr. Linda Partridge, University College London) (Wittmann et al., 2001 (link); Kerr et al., 2011 (link)), uas-human tandem Aβ42-22 amino acid linker-Aβ42 (gift from Dr. Damian Crowther, University of Cambridge) (Speretta et al., 2012 (link)), uas-GFP (gift from Dr. Mark Wu, Johns Hopkins University), uas-GCaMP6f [BL42747 (Shaw et al., 2018 (link))], uas-Ank2-RNAi line A (BL29438), uas-Ank2 RNAi (Vienna Drosophila RNAi Center stock number VDRC107238 and KK10497) and uas-Ank2 (gift from Dr. Ron R. Dubreuil, University of Illinois) (Mazock et al., 2010 (link)).

Higham J.P., Malik B.R., Buhl E., Dawson J.M., Ogier A.S., Lunnon K, & Hodge J.J. (2019). Alzheimer’s Disease Associated Genes Ankyrin and Tau Cause Shortened Lifespan and Memory Loss in Drosophila. Frontiers in Cellular Neuroscience, 13, 260.

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Drosophila Rearing and Genetic Lines

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Flies were raised on standard cornmeal food media at 25°C and 70% relative humidity under a 12:12-hour light: dark cycle. The “Cantonized” w¹¹¹⁸w (CS10) strain was used as a wild-type control. The MB-GAL80, UAS-GCaMP6m, and OK107-GAL4 flies were obtained from Bloomington Drosophila stock center. The UAS-eNpHR-YFP; UAS-eNpHR-YFP fly line was obtained from Ann-Shyn Chiang. The RNAi lines were obtained from the Vienna Drosophila RNAi Center or TRiP RNAi fly stocks. All RNAi lines from the Vienna Drosophila RNAi Center have been described previously[12 (link)]. The VT30604-GAL4, VT44966-GAL4, C739-GAL4, VT49246-GAL4, VT0765-GAL4, UAS-shi^ts, and tubP-GAL80^ts flies have been described[20 (link)].

Shyu W.H., Lee W.P., Chiang M.H., Chang C.C., Fu T.F., Chiang H.C., Wu T, & Wu C.L. (2019). Electrical synapses between mushroom body neurons are critical for consolidated memory retrieval in Drosophila. PLoS Genetics, 15(5), e1008153.

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Generating Drosophila Genetic Reporters

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WT flies used were Canton S (CS). Lar and sns mutants have been previously described: Lar^13.2, Lar⁴⁵¹, and sns^xb3 (gift of Dr. Susan Abmayr). The following lines were obtained from the Bloomington Stock Center: sns^Df, C155-GAL4, tubulin-GAL4, OK107-GAL4, sns^{MiMIC MI03001}, Lar^MiMIC02154, sns^EY08142, UAS-Lar RNAi (TRiP.HMS02186), UAS-Lar RNAi (TRiP.HMS00822), and UAS-Kirre RNAi (TRiP.HMC05791). Sns RNAi lines were from the Vienna Drosophila Resource Center: UAS-Sns RNAi (KK109442) and UAS-Sns RNAi (GD877). T2A-GAL4 lines were generated as described in Diao et al., 2015 (link). Briefly, flies carrying the MiMIC insertion were crossed with flies bearing the triplet ‘Trojan exon’ donor. The F₁ males from this cross carrying both genetic components were crossed to females carrying germline transgenic sources of Cre and ϕC31. The F₂ males from this cross that had all four genetic components were then crossed to a UAS-2xEGFP reporter line, and the resulting progeny were screened for T2A-GAL4 transformants.

Bali N., Lee H.K, & Zinn K. (2022). Sticks and Stones, a conserved cell surface ligand for the Type IIa RPTP Lar, regulates neural circuit wiring in Drosophila. eLife, 11, e71469.

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Drosophila Genetics: Fly Lines and Tools

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Flies were maintained on cornmeal-molasses medium at 25° unless otherwise stated. The collection of ENU-mutagenized Drosophila, including line 867 was a kind gift of Dr. Steven Robinow (University of Hawaii). UAS-brat, brat^ts1, brat^fs1 and brat^k06028 were obtained from Dr. Cheng-Yu Lee (University of Michigan). R9D11-mCD8-GFP was obtained from Dr. Jill Wildonger (University of Wisconsin-Madison). The following fly lines were obtained from the Bloomington Drosophila Stock Center at Indiana University: Df(2L)ED1272 (#24116), Df(2L)ED1203 (#8935), Df(2L)BSC341 (#24365), Df(2L)ED1231 (#9174), pcna-GFP (#25749), worniu-Gal4 (#56554), nSyb-Gal4 (#51635), CG15864^MB04166 (#24678), UAS-NICD (#52008), OK107-Gal4 (#854), UAS-mCD8-GFP (#5137), and insc-Gal4 (#8751). UAS-Brat-RNAi (#105054) was obtained from the Vienna Drosophila Resource Center (Dietzl et al. 2007 (link)).

Loewen C., Boekhoff-Falk G., Ganetzky B, & Chtarbanova S. (2018). A Novel Mutation in Brain Tumor Causes Both Neural Over-Proliferation and Neurodegeneration in Adult Drosophila. G3: Genes|Genomes|Genetics, 8(10), 3331-3346.

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Generating Transgenic Drosophila Lines Expressing Prion Proteins

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Flies expressing RaPrP-WT were described previously (Fernandez-Funez et al., 2010). The constructs carrying RaPrP-S174N, EqPrP-WT, EqPrP-SE167,168DQ, CaPrP-WT, CaPrP-D159N, and CaPrP-YD155,159DN cDNAs were synthesized at GenScript and cloned between EcoRI and NotI sites onto the pUAST Drosophila expression vector (Brand and Perrimon, 1993 (link)). The pUAST-based constructs were injected into yw embryos at Rainbow Transgenics following standard procedures (Rubin and Spradling, 1982 (link)) to generate multiple independent transgenic lines for each plasmid. The driver strains OK107-Gal4 (mushroom bodies), BG380-Gal4 (motor neurons), and da-Gal4 (ubiquitous), and the reporters UAS-LacZ and UAS-CD8-GFP were obtained from the Bloomington Drosophila Stock Center (http://fly.bio.indiana.edu). Fly stocks were maintained on standard Drosophila medium at 25°C. For experiments, homozygous females for the Gal4 strains were crossed with UAS males to generate progeny expressing PrP in the desired tissue. Crosses were placed at 25°C for two days, transferred to 28°C until the progeny completed development, and adults were aged at 28°C, unless otherwise indicated. All assays were performed using females.

Sanchez-Garcia J, & Fernandez-Funez P. (2018). D159 and S167 are protective residues in the prion protein from dog and horse, two prion-resistant animals. Neurobiology of disease, 119, 1-12.

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Drosophila Genetic Manipulation Toolkit

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Fly strains were maintained on a standard diet at 25 ℃, 12 light:12 dark cycles and 70% humidity. Actin-GeneSwitch (#9381), Dilp2-Gal4 (#37516), UAS-dSec16 RNAi (#53917), Ok107-Gal4 (#854), 104y-Gal4 (#81014), Dsk-Gal4 (#51981), Dh44-Gal4 (#51987), Lk-Gal4 (#51992), OK371-Gal4 (#26160), Chat-Gal4 (#6793), 55D01-Gal4 (#39110), Ppk-Gal4 (#79278) and Pdf-Gal4 (#6899) were purchased from the Bloomington Stock Center.

Zhang R.X., Li S.S., Li A.Q., Liu Z.Y., Neely G.G, & Wang Q.P. (2022). dSec16 Acting in Insulin-like Peptide Producing Cells Controls Energy Homeostasis in Drosophila. Life, 13(1), 81.

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Genetics of Drosophila Learning Behavior

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The following strains were used: y,w and CS were used as the control; y, w, dlg^XI-2; y, w, dlgS97⁵; y, w, dlgA^40.2; (Bloomington Stock Center, Bloomington, IN, USA). All of the strains were backcrossed six-fold with Canton-S to standardize the genetic background (Cantonization). For the adult associative olfactory learning experiments, the strains used were: W(CS); Ok107-GAL4 (kind gift from Dr. Jorge Campusano); UAS-RNAi-dlg (Vienna Drosophila Resource Center, AT); UAS-dsRNA-dlg1; Orco-GAL4; GH146-GAL4; Tenm-GAL4;117y-GAL4; C305A-GAL4; Elav-GAL4 (Bloomington Stock Center, Bloomington, IN); Ok107-GAL4; UAS-DlgS97 (Kind gift from Dr. U. Thomas). For the dNR1 and dNR2 overexpression, UAS-GFP-CD8-dNR1 and UAS-GFP-CD8-dNR2 flies were used. All of the flies were grown at 22 °C.

Bertin F., Moya-Alvarado G., Quiroz-Manríquez E., Ibacache A., Köhler-Solis A., Oliva C, & Sierralta J. (2022). Dlg Is Required for Short-Term Memory and Interacts with NMDAR in the Drosophila Brain. International Journal of Molecular Sciences, 23(16), 9187.

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Genetic Manipulation of Drosophila Cytoskeleton

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Flies were raised at 25 °C under standard conditions. The following mutant strains were used: w¹¹¹⁸ (BL #3605), Elav-Gal4 (BL #8760 and 8765), OK107-Gal4 (BL #854) provided by the Bloomington Drosophila Stock center, dDAAM^Ex4 [28 (link)], frl⁵⁹ [29 (link)], DAAM^Ex4; frl⁵⁹/TM3, twi-Gal4, UAS-EGFP, UAS-DAAM [30 (link)], UAS-DAAM^I732A [28 (link)], UAS-DAAM^K881A [31 (link)], UAS-DADG [28 (link)] and UAS-CDAAM [30 (link)]. Where necessary, zygotic mutants were selected by using a CyO, twi-Gal4, UAS-EGFP or TM3, twi-Gal4, UAS-EGFP balancer chromosome. For live imaging, the ElavGal4,EB1::GFP line was crossed to UAS-CDAAM or UAS-CDAAM^I732A. The UAS-DAAM^FH2R/K-A, UAS-CDAAM^I732A and UAS-CDAAM^FH2R/K-A transgenic constructs were generated as described below. Drosophila transgenesis was carried out by using the PhiC31 integrase and all transgenes were integrated into the attP40 landing site on the second chromosome.

Földi I., Tóth K., Gombos R., Gaszler P., Görög P., Zygouras I., Bugyi B, & Mihály J. (2022). Molecular Dissection of DAAM Function during Axon Growth in Drosophila Embryonic Neurons. Cells, 11(9), 1487.

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Drosophila Transgenic RNAi Lines

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The following fly stocks were used in this study: UAS-H1KD RNAi lines (TH00868.N, TH00825.N, and TH00826.N), a UAS-mof RNAi line (TH00870.N), UAS-dBigH1 RNAi lines (TH11322.N and TH11323.N), UAS-GFP RNAi line (TH00782.N), UAS-GFP (TH10512.N), P[nosP-GAL4::vp16]46 (link), y, hs-FLP; nos<STOP<GAL4::VP16, UASp-GFP/CyO29 (link), P[bamP-GFP]22 (link), Dad-LacZ35 (link), bam^Δ86 (ref. 47 (link)), UAS-p35 (ref. 48 (link)), 1824-GAL4 (AB1-GAL4; Bloomington #1824), OK107-GAL4 (w*;;;P{GawB}ey^OK107, a gift from Dr Yi Zhong) and c587-Gal4 (Bloomington #25421).
Sense Oligos of the transgenic RNAi lines are:
TH00782.N: 5′-CCCGAAGGTTATGTACAGGAA-3′
TH00825.N: 5′-AAGCAAGAAGGTAGCCTCTAA-3′
TH00826.N: 5′- TAGCGAAAGCGTCAAAGGCAA-3′
TH00868.N: 5′-ACCAGCGACAGTTGAGAAGAA-3′
TH00870.N: 5′-CTCGACCTCAGCGGTGTCCAA-3′
TH11322.N: 5′-CGGCGAAGTGGTGATGGTTAA-3′
TH11323.N: 5′-ATGGTTAAGCGATCCTTTAAA-3′.
All Drosophila stocks were maintained at 25 °C with 60% humidity on standard cornmeal/sugar/agar media unless otherwise specified.

Sun J., Wei H.M., Xu J., Chang J.F., Yang Z., Ren X., Lv W.W., Liu L.P., Pan L.X., Wang X., Qiao H.H., Zhu B., Ji J.Y., Yan D., Xie T., Sun F.L, & Ni J.Q. (2015). Histone H1-mediated epigenetic regulation controls germline stem cell self-renewal by modulating H4K16 acetylation. Nature Communications, 6, 8856.

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