W1118

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W1118 is a genetically modified Drosophila melanogaster (fruit fly) strain that is commonly used in research laboratories. It serves as a reference strain and is widely employed as a genetic background for various experiments and transgenic manipulations.

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22 protocols using w1118

Drosophila Rearing and Genetic Manipulation

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Flies were reared on standard cornmeal food (130 g of yarn agar, 248 g of Baker’s yeast, 1223 g of cornmeal, and 1.5 liters of sugar beet syrup in 20 liters of distilled water) and kept in a 25°C incubator with light-dark cycle. Fly stocks were obtained from the Bloomington Drosophila Stock Center (BDSC) and the Vienna Drosophila Resource Center. For stainings and SING assays, 2- to 5-day-old female flies were kept on control diet or on filter paper soaked with 10% sucrose solution and 50 or 100 μM rotenone or 20 μM CsA. We obtained the UAS-CaMPARI2 and the TH-Gal4 line from A. Schoofs and the UAS mito-ER tether lines from E. Ziviani. Other fly lines used in this study were w¹¹¹⁸ (BDSC, no. 6326), elav-Gal4 (BDSC, no. 8760), tubulin-Gal4 (BDSC, no. 5138), mef-Gal4 (BDSC, no. 27390), handC-Gal4 (37 (link)), actin-Gal4 (BDSC, no. 4414), armadillo-Gal4 (BDSC, no. 1561), UAS-Creld-HA (this study), UAS-mCherry-mitoOMM (BDSC, no. 66533), Drp1-HA (BDSC, no. 42208), UAS Pink1.C (BDSC, no. 51648), UAS catalase-RNAi (BDSC, no. 34020), UAS Duox-RNAi (BDSC, no. 33975), UAS-Sod1 (BDSC, no. 24750), UAS ND-39 (BDSC, no. 15821), UAS GFP-KDEL (BDSC, no. 9899), Pink1⁵ (BDSC, no. 51649), and park²⁵ (52 (link)).

Paradis M., Kucharowski N., Edwards Faret G., Maya Palacios S.J., Meyer C., Stümpges B., Jamitzky I., Kalinowski J., Thiele C., Bauer R., Paululat A., Sellin J, & Bülow M.H. (2022). The ER protein Creld regulates ER-mitochondria contact dynamics and respiratory complex 1 activity. Science Advances, 8(29), eabo0155.

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Fly Gut Stem Cell Regulation

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Fly stocks were maintained at 25°C on standard food under a ∼12 h/12 h light/dark cycle. Food consisted of 15.8g yeast, 9g soy flour, 5.2g agar, 67g cornmeal, and 0.5% propionic acid. To avoid larval overpopulation, <30 adult flies per vial were transferred to new food vials every 2–3 days.
The following stocks were used in this study: esg-Gal4,tub-Gal80ts,UAS-GFP/CyO (esg^ts>GFP (gift from Bruce Edgar) [48 (link)]; FRT82B, tub-Gal80/TM6B (gift from Bruce Edgar) [49 (link)]; Myo1A-Gal4,tub-Gal80ts,UAS-GFP/CyO (Myo1A^ts> GFP) (gift from Bruce Edgar) [48 (link)]; UAS-CncC (gift from Dirk Bohmann) [50 (link)]; UAS-Png1WT; UAS-Png1C303A; Png1ex18; Png1ex14 (gift from Hamed Jafar-Nejad) [28 (link)]; UAS-OGARNAi (VDRC, #106670); UAS-OGARNAi (VDRC, #41822); UAS-OGTRNAi (VDRC, #18610); UAS-OGTRNAi (VDRC, #18611); UAS-Png1RNAi (VDRC, #103607); w¹¹¹⁸ (BDSC, #3605); UAS-Png1RNAi (BDSC, #54853); UAS-ENGase RNAi (BDSC, #64609).
The UAS-OGT line that contains Myc epitope tag in the N terminus (UAS-Myc-OGT) was made by P-element-mediated transformation and OGAdel.1 mutant was generated by standard P-element excision [51 (link)].
For transgene expression at specific developmental stages, the Gal80ts technique was used. The flies were set up and maintained at 22°C until adulthood. After maintaining the flies at 29°C, the midguts were dissected and analyzed.

Na H.J., Abramowitz L.K, & Hanover J.A. (2022). Cytosolic O-GlcNAcylation and PNG1 maintain Drosophila gut homeostasis by regulating proliferation and apoptosis. PLoS Genetics, 18(3), e1010128.

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Generating Transgenic Drosophila Fly Lines

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The following fly lines are available at the Bloomington Drosophila Stock Center (Bloomington, IN): w¹¹¹⁸, Df(3L)BSC392 (BDSC-24416), 24B-Gal4, elav^c155-Gal4, Act-Gal4, hs-Cre and vasa-Cas9. UAS-FLAG-SH3PX1 flies were generated by cloning the full-length SH3PX1 cDNA into pBID-UASC-FG (Wang et al., 2012 (link)) and integrating into the attP2 landing site (Groth et al., 2004 (link)).

Ukken F.P., Bruckner J.J., Weir K.L., Hope S.J., Sison S.L., Birschbach R.M., Hicks L., Taylor K.L., Dent E.W., Gonsalvez G.B, & O'Connor-Giles K.M. (2016). BAR-SH3 sorting nexins are conserved interacting proteins of Nervous wreck that organize synapses and promote neurotransmission. Journal of Cell Science, 129(1), 166-177.

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Drosophila Genetics: Fly Stock Maintenance

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Unless otherwise indicated, we maintained all fly stocks at 25 °C and 60% relative humidity under a 12 h:12 h light:dark cycle on standard cornmeal-yeast-corn syrup medium with 1.5 g/L of the anti-fungal Tegosept. We acquired the following fly stocks from either the Bloomington Drosophila Stock Center (BDSC) or the Vienna Drosophila Resource Center (VDRC) for our experiments: Canton-S (BDSC #1), w¹¹¹⁸ (BDSC #5905), NPF-GAL4 (BDSC #25681, #25682), UAS-myr::GFP (BDSC #32197, #32198), UAS-NPF-IR (VDRC #108772), 386Y-GAL4 (BDSC #25410), NPFR^c01896 (BDSC #10747), NPFR^Def (BDSC #1982), Orco-GAL4 (BDSC #23292), Or22a-GAL4 (BDSC #9951), UAS-NPFR-IR (VDRC #9605). We generated the UAS-NPFR-RD and UAS-NPFR-RB stocks by cloning NPFR cDNAs from Drosophila heads, subcloning them into a pUAS-T attB-containing vector, and injecting the resulting plasmids into eggs of a fly stock containing an attP landing site (BDSC #9736) using standard techniques.

Lee S., Kim Y.J, & Jones W.D. (2017). Central peptidergic modulation of peripheral olfactory responses. BMC Biology, 15, 35.

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Drosophila Genetics Resource Database

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All fly stocks were maintained at 25°C on standard medium unless otherwise stated. The following fly lines were used in this study: UAS-sna⁺ (FlyORF F000066), UAS-esg⁺ (FlyORFF000254), UAS-wor⁺ (FlyORF F000155), UAS-sna^RNAi (BDSC BL28679), UAS-esg^RNAi (BDSC BL42846), UAS-wor^RNAi (VDRC v6248), UAS-lsd1^RNAi (BDSC BL65020), UAS-lsd2^RNAi (BDSC BL32846), UAS-lsd1⁺ (BDSC BL31784), UAS-lsd2⁺ (BDSC BL19733), Ppl-Gal4 (BDSC BL58768), R4-Gal4 (BDSC BL 33832), caudal (Cd)-Gal4 (BDSC BL3042), w1118 (BDSC BL 6326), and tPGH (BDSC BL 8164). Tubulin-Gal80^ts (BDSC BL7180). Hand-Gal4 is a gift from Lauren Perrin (AMU, France), and Myo1A-Gal4 is a gift from Bruce Edgar (University of Utah).

Liu Y., Bao H., Wang W, & Lim H.Y. (2019). Cardiac Snail family of transcription factors directs systemic lipid metabolism in Drosophila. PLoS Genetics, 15(11), e1008487.

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Drosophila Stocks Maintenance and Characterization

Cited 1 time

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Drosophila stocks were maintained on standard fly food (25 g/L corn flour, 5 g/L lyophilized agar, 50 g/L sugar, 50 g/L fresh yeast, 2.5 mL/L Tegosept [10% in ethanol], and 2.5 mL/L propionic acid) at 25 °C in a 12 h light/dark cycle. All experiments were performed in the same standard conditions, otherwise differently specified. The following fly strains were purchased from the Bloomington Drosophila Stock Center (BDSC, Indiana University, Bloomington, IN, USA): w¹¹¹⁸ (BDSC #3605); elav-GS (BDSC #43642); UAS-TBPH (BDSC #93601); UAS-TBPH.F-L (BDSC #93781); UAS-mCD8-GFP (BDSC #30002); Su(var)3-9¹/TM3 (BDSC #6209); UAS-Su(var)3-9.lacI (BDSC #93147); UAS-hSUV39H1.HA (BDSC #84799); elav-GAL4 (BDSC #77894); egg¹⁴⁷³/SM1 (BDSC #30565). The Su(var)3-9⁶/TM6B allele was a kind gift of Gunter Reuter [49 (link)], the G9a^RG5 allele was a kind gift of Marion Delattre [53 (link)].

Marzullo M., Romano G., Pellacani C., Riccardi F., Ciapponi L, & Feiguin F. (2023). Su(var)3-9 mediates age-dependent increase in H3K9 methylation on TDP-43 promoter triggering neurodegeneration. Cell Death Discovery, 9, 357.

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Drosophila Genetic Toolkit: Strains and Reagents

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Misexpression P{Mae-UAS.6.11} inserts (LA lines) were obtained from John Merriam, UCLA (Los Angeles, California). UAS-RNAi lines were obtained from the Vienna Drosophila RNAi Center (VDRC, Vienna, Austria), the National Institute of Genetics (NIG, Kyoto, Japan), and the Bloomington Drosophila Stock Center (TRiP lines, BDSC, Bloomington, Indiana). The lines UAS-FLP.JD1, UAS-2XEGFP, P{GAL4-Act5C(FRT.CD2).P}S, UAS-human Raf^ACT, Df(3R)mbc-R1, UAS-RpS8^PD01446, and w¹¹¹⁸ (BDSC 5905) were from the BDSC. Pvr^c02195 was from Exelixis (available from BDSC, obtained from D Montell). Hml^Δ-gal4 UAS-2XEGFP (S Sinenko), Antp-gal4/TM6B Tb (S Cohen), P{ubi-gal80 ts}10; Antp-gal4/TM6B Tb (this lab), domeless-gal4 UAS-2XEYFP/FM7i (this lab), UAS-DAlk^ACT (R Palmer), dome-MESO-lacZ (S Brown), Pxn-gal4 (M Galko), UAS-STAT^ACT (E Bach), UAS-ADGF-A (T Dolezal), collier¹; P(col5-cDNA)/CyO-TM6B, Tb (M Crozatier), srpHemo-gal4 (K Brückner), and Hand-gal4 (Z Han) have been previously described.

Mondal B.C., Shim J., Evans C.J, & Banerjee U. (2014). Pvr expression regulators in equilibrium signal control and maintenance of Drosophila blood progenitors. eLife, 3, e03626.

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Fly Strain Protocols for Drosophila Research

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The following fly strains were used: drpr^∆5FRT2A^{40 (link),41 (link)}, UAS- Relish RNAi (Vienna Drosophila Stock Center #49414), AttGFP (^{26 (link)} provided by Dr. Mitch Dushay), repo-GAL4/TM3 (Bloomington Drosophila Stock Center (BDSC) #7415), elav-GAL4 (BDSC #458), ppl-GAL4 (^{42 (link)} RRID:BDSC_58768, provided by Dr. Michael O’Connor) and w¹¹¹⁸ (BDSC). All fly lines were raised at 25°C on standard cornmeal and molasses food.

Elguero J.E., Liu G., Tiemeyer K., Gandevia H., Duro L, & McCall K. (2023). Defective phagocytosis leads to neurodegeneration through systemic increased innate immune signaling. bioRxiv.

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Drosophila Gametogenesis Genetic Toolkit

Cited 3 times

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All fly stocks were raised on standard Bloomington medium at 25°C. lky RNAi flies were placed at 29°C after eclosion for desired days before dissection. For heat-shock mediated induction of RNAi in aged (day 21) vs. young (day 0–1) flies, hs-FLP; nos-FRT-stop-FRT-Gal4 [59 (link)] was crossed with RNAi lines and progenies were heat-shocked at 37˚C for 60 minutes for 2 times. Then, testes were dissected at the indicated days after the heat shock. Flies cultured for aging were placed into fresh food vials every 48 hours and dissected at indicated ages. Temperature shift was performed by culturing flies at room temperature and shifted to 29˚C upon eclosion for the 4 days before analysis. Combinations of Tub-Gal80^ts (a gift from Cheng-Yu-Lee) with c587Gal4 (a gift from Yukiko M. Yamashita) were used. The following fly stocks were used:: αTub84B^K40A (FBal0345033, [52 (link)] αTub84B knock-in, gift from J. Wildonger), dTAT^KO (FBal0345149, [15 (link)], gift from J. Wildonger), UAS-nosTub3’UTR, nosGal4VP16, tjGal4 (gifts from Yukiko M. Yamashita), nos^L7 and nos^BN (gifts from R. Lehmann). The following stocks were obtained from the Bloomington stock center, w1118 (BDSC 3605), yw (BDSC 1495), lky deficiency, Df(1)BSC586 (BDSC 25420) and UASp-hts.mCherry (BDSC 66171).

Antel M., Simao T., Bener M.B, & Inaba M. (2022). Drosophila CG17003/leaky (lky) is required for microtubule acetylation in early germ cells in Drosophila ovary. PLOS ONE, 17(11), e0276704.

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Drosophila Genetic Toolkit: Diverse Strains

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All fly stocks were maintained at 25 °C on standard medium unless otherwise stated. The following stains were used in this study: w¹¹¹⁸ (BDSC BL6326), Act-Gal4 (BDSC BL3954), Da-Gal4 (BDSC BL55851), Ub-Gal4 (BDSC BL32551), Caudal (Cad)-Gal4 (BDSC BL3042), R4-Gal4 (BDSC BL33832), ElaV-Gal4 (BDSC BL458), Mhc-Gal4 (BDSC BL38464), Hsp70-Gal4 (BDSC BL2077), Escargot-Gal4 (Kyoto DGRC 114-042), Prospero-Gal4 (BDSC 80572), UAS-dSLC5A5^RNAi (BDSC BL63568), UAS-dSLC5A5^RNAi (VDRC 40650), and UAS-shibire^RNAi (BDSC BL28513). The MyoIA-Gal4 fly line is a gift from Dr. Bruce Edgar (University of Utah, Salt Lake City, UT, USA). The dSLC5A5-FLAG clone (UFO11244) was obtained from the Drosophila Genomics Resource Center (DGRC) and contains full-length dSLC5A5 cDNA with a C-terminus FLAG HA tag in a pUAST vector. UAS-dSLC5A5-FLAG transgenic flies were generated by BestGene Inc. (Chino Hills, CA, USA). For each cross, the Gal4 or UAS-RNAi progenies derived from the cross are used as controls.

Li Y., Wang W, & Lim H.Y. (2021). Drosophila Solute Carrier 5A5 Regulates Systemic Glucose Homeostasis by Mediating Glucose Absorption in the Midgut. International Journal of Molecular Sciences, 22(22), 12424.

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