Fly stocks were maintained on cornmeal/agar/yeast food at 21°C, except where noted. Before immunofluorescence staining, newly eclosed flies were fed wet yeast paste every day for 2–4 days. Unless specified, yw (BDSC 1495) was used as the control. The following stocks were obtained from the Bloomington Drosophila Stock Center: c355 GAL4 (BDSC 3750), actin-5C GAL4 (BDSC 8807), mgst1KG04713 (BDSC 13839), Su(P)EY13245 (BDSC 20866), p23EY05607(BDSC 16661), akr1BPL00034 (BDSC 19594), akr1BEY07011 (BDSC 16777), p23 RNAi HMJ24151 (BDSC 62911), p23 RNAi-2 GL01292 (BDSC 41862), akr1B deficiency-1 DF(3L)BSC577 (BDSC 25411), akr1B deficiency-2 DF(3L)ED4475 (BDSC 8069), akr1B RNAi HMS05657 (BDSC 67838), akr1B RNAi-2 HMC05226 (BDSC 62219) and UAS Dicer 2 (BDSC 24651). The following stocks were obtained from the Exelexis Stock Center: mgst1d10243, akr1Bd00405 and pxtf01000 (Thibault et al., 2004 (link)). The oskar GAL4 line (second chromosome; BDSC 44241) was a generous gift from Anne Ephrussi [European Molecular Biology Laboratory; (Telley et al., 2012 (link))]. Expression of the RNAi lines were achieved by crossing to actin-5C GAL4, c355 GAL4 or oskar GAL4, maintaining fly crosses at 21°C and maintaining progeny at 29°C for 5–6 days. UAS Dicer was used in combination with c355 to enhance RNAi efficiency where noted in the figure legends.
Uas dicer2
Manufactured by BD
Sourced in United States, Austria
The UAS-dicer2 is a laboratory equipment designed for processing biological samples. It functions as a cutting and shredding device for tissue and other materials. The core purpose of the UAS-dicer2 is to prepare samples for further analysis or processing, without additional interpretation or intended use.
Automatically generated - may contain errors
Lab products found in correlation
C355 gal4, by BD (2 mentions)
Mgst1kg04713, by BD (2 mentions)
Su p ey13245, by BD (2 mentions)
P23ey05607, by BD (2 mentions)
Akr1bpl00034, by BD (2 mentions)
Akr1bey07011, by BD (2 mentions)
P23 rnai hmj24151, by BD (2 mentions)
P23 rnai 2 gl01292, by BD (2 mentions)
Actin5c gal4, by BD (1 mentions)
Ppk gal4, by BD (1 mentions)
Uas khc rnai, by BD (1 mentions)
C306 gal4, by BD (1 mentions)
Pnr gal4, by BD (1 mentions)
Uas p35, by BD (1 mentions)
Quikchange lightning site directed mutagenesis kit, by Agilent Technologies (1 mentions)
W1118, by Bloomington Drosophila Stock Center (1 mentions)
Uas dicer2, by Vienna Drosophila Resource Center (1 mentions)
Uas tkvca, by BD (1 mentions)
Uas rab11 gfp, by BD (1 mentions)
Uas mcherrycaax, by BD (1 mentions)
7 protocols using uas dicer2
1
Drosophila Genetics and Immunofluorescence
2
Drosophila Kinesin Heavy Chain Alleles
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The Khc alleles Khc27 (null allele), Khc22 (amino acid replacement P945S), and Khc23 (amino acid replacement E164K) were provided by W. Saxton, University of California, Santa Cruz, Santa Cruz, CA (Brendza et al., 1999 (link)). UAS-Khc::BFP was provided by V. Gelfand (Northwestern University, Chicago, IL; Winding et al., 2016 (link)). The following fly strains were obtained from the BDSC and the Vienna Drosophila Resource Center: UAS-hTfR::GFP (BDSC stock no. 36858); ppk-CD4::Tomato (BDSC stock no. 35845); ppk-CD4::GFP (BDSC stock no. 35843); ppk-Gal4 (BDSC stock nos. 32078 and 32079); ensconsin alleles, including enscΔC, which lacks the majority of coding exons (Sung et al., 2008 (link); BDSC stock no. 51318), and Df(3L)enscΔ3277, a deficiency that removes ensconsin (BDSC stock no. 51319); UAS-Khc-RNAi (BDSC stock no. 35770); UAS-dicer2 (BDSC stock no. 24650); and UAS-dlic-RNAi (stock no. 41686; Vienna Drosophila Stock Center). All transgenic markers (e.g., ppk-ManII::GFP) were used as a single copy. Khc mutants were analyzed in trans to the null allele Khc27 unless otherwise noted.
3
Drosophila Genetics: Protocols and Stock Utilization
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Fly stocks were maintained on cornmeal/agar/yeast food at 21°C, except where noted. Before immunofluorescence staining, newly eclosed flies were fed wet yeast paste every day for 2–4 days. Unless specified, yw (BDSC 1495) was used as the control. The following stocks were obtained from the Bloomington Drosophila Stock Center: c355 GAL4 (BDSC 3750), c306 GAL4 (BDSC 3743), mgst1KG04713 (BDSC 13839), Su(P)EY13245 (BDSC 20866), p23EY05607(BDSC 16661), akr1BPL00034 (BDSC 19594), akr1BEY07011 (BDSC 16777), p23 RNAi HMJ24151 (BDSC 62911), p23 RNAi-2 GL01292 (BDSC 41862), akr1B RNAi HMS05657 (BDSC 67838), akr1B RNAi-2 HMC05226 (BDSC 62219) and UAS Dicer 2 (BDSC 24651). The following stocks were obtained from the Exelexis Stock Center: mgst1d10243, akr1Bd00405 and pxtf01000 (Thibault et al., 2004 (link)). The oskar GAL4 line (second chromosome; BDSC 44241) was a generous gift from Anne Ephrussi (European Molecular Biology Laboratory; (Telley et al., 2012 (link))). Expression of the RNAi lines were achieved by crossing to c355 GAL4 or oskar GAL4, maintaining fly crosses at 21°C and maintaining progeny at 29°C for 5–6 days. UAS Dicer was used in combination with c355 to enhance RNAi efficiency were noted in the figure legends.
4
Drosophila Genetic Toolkit for Development
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The following fly stocks were used in this study: w1118 (BDSC#3601) as WT flies, y w hep1 and y w hepr75 [1 (link)], UAS-hepact (BDSC#9306), pucE69 (puc-Z)[54 (link)], 69B-GAL4 (BDSC#1774), pnr-GAL4 (BDSC#3039), en-GAL4 (gift from A. Brand), AS-GAL4 (c381-GAL4; BDSC#3734), enX31 [55 (link)], UAS-en [56 (link)], UAS-HA::VP16::en [39 (link)], Scr17 (BDSC#3400), Antp[Ns-rvC4] (BDSC#1830), Ubx1 (BDSC#626), abdAM1 and AbdBD18 [57 (link)], UAS-Scr::HA, UAS-Ubx::HA and UAS-abdA::HA [58 (link)], UAS-Antp [59 (link)], UAS-AbdBm (BDSC#913), UAS-AbdBi (VDRC#12024)[60 (link)], UAS-Dicer2 (BDSC#24650), hthP2 [61 (link)], TM3,dfd-lacZ [62 (link)]. For live imaging, we used arm::GFP (BDSC#8556 on chromosome II; BDSC#8555 on chromosome III) and created the recombinant line arm::GFP,Abd-BD18 that we balanced with TM3,twi-GAL4,UAS-GFP (BDSC#6663) to select the homozygote mutant embryos. We also created the following line for the Abd-B RNAi: UAS-Dicer2;UAS-CD8::RFP,UAS-Abd-Bi, allowing the selection of AbdBi embryos thanks to the expression of CD8::RFP.
5
Genetic Manipulation of Drosophila for Research
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Drosophila stocks were maintained on a standard cornmeal medium. The following transgenic stocks were utilized in this study: P{w+; UAS-GFP::dPRLWT}, P{w+; UAS-GFP::dPRLC173S}, P{w+; GMR-RGH-miRNA} [34 (link)], P{w+; GMR-reaper-miRNA} [34 (link)], P{w+; GMR-grim-miRNA} [34 (link)], P{w+; GMR-hid-miRNA} [34 (link)], P{w+; UAS-dPRL-IR45518} (Vienna Drosophila Resource Center (VDRC, Vienna, Austria, stock No. 45518), GMR-Gal4 (Bloomington Drosophila Stock Center (BDSC, Bloomington, IN, USA, stock No. 1104), UAS-dicer2 (BDSC, stock No. 24646), P{w+; UAS-p35} (BDSC, stock No. 5072). To ensure the GAL4 expression and efficiency for RNA interference, flies were raised at 27 °C for pupal dissection. The wild-type coding sequence of dPRL were subcloned into the pUAST vector containing a GFP coding sequence upstream of the cloning site for transgenes. The QuikChange Lightning Site-Directed Mutagenesis kit (Agilent Technologies, Santa Clara, CA, USA) was used to generate the Cys173 to Ser substitution in the dPRL coding sequence. Transgenic flies were generated according to the standard protocol via microinjection.
6
Drosophila Strain Characterization and Manipulation
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Several Drosophila strains were used, including: w1118 (Bloomington, USA, BDSC); witA12 (BDSC); C380-GAL4 (BDSC); OK6-GAL4 (BDSC), UAS-dicer2; UAS-dsRNA-atl (Vienna Drosophila Resource Center, Austria, VDRC), UAS-dicer2, UAS-tkv-CA, UAS-rab4-GFP, UAS-rab11-GFP and Rab7-YFP (BDSC). A detailed table with all the stocks used can be found in Additional file 1 : Table S1.
7
Imaging Drosophila Cytoskeleton and Lineage
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The following Drosophila stocks were used: R10H12-gal4 (Pfeiffer et al., 2008 (link); BDSC 48278), UAS-mCherryCAAX (BDSC 59021), UAS-mCherryNLS (BDSC 38425), enhancer trap lines sr-gal4md710 (Usui et al., 2004 (link), BDSC 2663), UAS-Lifeact.GFP (BDSC 35544), UAS-dar1RNAi (BDCS 31987), dar13010 (BDSC 65269), UAS-GC3Ai (Schott et al., 2017 (link), BDSC 84346), UAS-serpRNAi (BDSC 63556), UAS-vermRNAi (BDSC 57188), UAS-lolalRNAi (BDSC 35722), UAS-Dicer2 (BDSC 24650), UAS-SrDN (gift from Vorbrüggen and Jäckle, 1997 (link)) and G-TRACE line (Evans et al., 2009 (link), BDSC 28280).
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