Uas dicer

Manufactured by Bloomington Drosophila Stock Center

Sourced in Spain

The UAS-dicer is a genetic tool used in Drosophila research. It functions as a double-stranded RNA (dsRNA) expression system, which can be used to induce RNA interference (RNAi) and knockdown the expression of target genes in Drosophila.

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4 protocols using uas dicer

Genetic Tools for Drosophila Muscle Studies

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Strain y¹w¹¹⁸ was used as control. The following stocks were used: tub-GAL80ts (7108), UAS-Dicer (24651) and UAS-perd RNAi JF01159 (31584) from Bloomington Drosophila Stock Center (http://flystocks.bio.indiana.edu/). Kettin-GFP (ZCL2144) from FlyTrap (http://flytrap.med.yale.edu/). UAS-perd RNAi (106680) and UAS-mys RNAi (29619) from Vienna Drosophila RNAi Center (http://stockcenter.vdrc.at). Zasp-GFP (110740) from Drosophila Genetic Resource Center (http://www.dgrc.kit.ac.jp/). Mef2-GAL4 (Ranganayakulu et al., 1996 (link)). UAS-mCD8-GFP (Lee and Luo, 1999 (link)). UAS-rhea-mCherry (Venken et al., 2011 (link)). UAS-kon-HA (Schnorrer et al., 2007 (link)). 1151-GAL4 was a gift from Lingadahalli S. Shashidhara (Centre for Celular and Molecular Biology, Hyderabad, India). For transient expression assays, the different UAS lines were crossed with tub-GAL80ts; Mef2-GAL4 at 18°C, and the UAS expression was induced switching to 29°C. The temperature shift was made when larvae were between the first and the second larval stage, unless otherwise specified.

Pérez-Moreno J.J., Bischoff M., Martín-Bermudo M.D, & Estrada B. (2014). The conserved transmembrane proteoglycan Perdido/Kon-tiki is essential for myofibrillogenesis and sarcomeric structure in Drosophila. Journal of Cell Science, 127(14), 3162-3173.

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Drosophila Neurodegeneration Genetics

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All fly stocks were maintained and raised under standard conditions at 25°C. Canton S was used as wild type. UAS-SWS^R133A and UAS-PKA-C3 are described in [24] (link), and sws¹ in [19] (link). elav-GAL4, UAS-dicer, and UAS-lacZ were obtained from the Bloomington stock center and Appl-GAL4 was kindly provided by L. Torroja (Universidad Autonoma de Madrid, Spain). The PKA-C3 RNAi line (Pka-C3^NIG.6117R) was obtained from the National Institute of Genetics (NIG-Fly), Japan.

Wentzell J.S., Cassar M, & Kretzschmar D. (2014). Organophosphate-Induced Changes in the PKA Regulatory Function of Swiss Cheese/NTE Lead to Behavioral Deficits and Neurodegeneration. PLoS ONE, 9(2), e87526.

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Genetic Tools for Neuronal Manipulation

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sss^P1, sss-Gal4 and UAS-sss flies were described previously [6 (link), 7 (link)]. Other lines were provided as follows: Sh^mns (Chiara Cirelli), UAS-shi^ts (Toshi Kitamoto), UAS-TRPA1 (Paul Garrity). UAS-Dα3 (stock d03389) was from the Exelixis collection at Harvard. Other fly lines were obtained from the Bloomington Stock Center (UAS-dicer [24650]; UAS-tdTomato [32221]; UAS-nAChR RNAi's [28688, 27493, 27671, 31985, 25943, 25835, 27251, 31883, 28038 and 25927]). UAS-lynx1 flies were generated by targeting UAS-lynx1 in pUAST-attB to the attP site of y¹,w^67c23;;attP2 flies (Rainbow Transgenics, Camarillo, CA) and outcrossing transgenic animals into a w¹¹¹⁸ iso31 background for 2 generations.

Wu M., Robinson J.E, & Joiner W.J. (2014). SLEEPLESS is a bi-functional regulator of excitability and cholinergic synaptic transmission. Current biology : CB, 24(6), 621-629.

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Genetic Manipulation Protocols for Insect Species

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Wild-type T. castaneum strain and the enhancer-trap line pu11 (obtained from Y. Tomoyasu, Miami University, Oxford, Ohio) were reared on organic wheat flour containing 5% nutritional yeast, and maintained at 29°C in constant darkness. Flies were raised on standard D. melanogaster medium at 25°C, unless otherwise required. Oregon R flies (OR-R, used as a wild type control), ActGAL4, rnGal4, CiGAL4, UASGFP and UASdicer (used to enhance RNAi effectiveness) were obtained from the Bloomington Stock Center (BDSC). UAS-DE93^RNAi (KK108140; GD4449), and UAS-DKr-h1^RNAi (KK107935; GD51282) are from the Vienna Drosophila RNAi Center (VDCR). For clonal analysis, hsflp;Tub>y>Gal4;UASGFP females were crossed with males carrying UASKr-h1^RNAi. Embryos were kept at 25°C until late L2, incubated 1 hour at 37°C and transferred to 25°C until late L3. B. germanica specimens were reared in the dark at 30 ± 1°C and 60–70% relative humidity.

Ureña E., Chafino S., Manjón C., Franch-Marro X, & Martín D. (2016). The Occurrence of the Holometabolous Pupal Stage Requires the Interaction between E93, Krüppel-Homolog 1 and Broad-Complex. PLoS Genetics, 12(5), e1006020.

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