Uas nlsgfp

Manufactured by Bloomington Drosophila Stock Center

The UAS-nlsGFP is a genetic construct that expresses a nuclear-localized GFP (green fluorescent protein) reporter under the control of the upstream activating sequence (UAS). It is a commonly used tool in Drosophila genetics for visualizing the expression and localization of target genes in a tissue-specific manner.

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6 protocols using uas nlsgfp

Drosophila Genetic Manipulation Protocols

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Standard methods were used for propagating fly stocks. For all experiments, embryos and larvae were raised at 25 °C, unless otherwise noted. The following lines were used: CQ2-GAL4 (Bloomington stock center [BL] 7468), OK6-GAL4 (BL 64199), hsFLP; UAS(FRT.stop)myr::smGdP-HA, UAS(FRT.stop)myr::smGdP-V5-THS UAS(FRT.stop)myr::smGdP-FLAG (BL 64085), UAS-myr-GFP (BL 32198), UAS-nls-GFP (BL 6452), UAS-Hb; UAS-HB/TM2 (BL 8504), w1118 (BL 36005), MHC-CD8-GCaMP6f-Sh (BL 67739) ac:VP16, gsb:v8v (aka NB7-1-GAL4, gift of M. Kohwi), VGlut-lexA/cyo (BL 60314), lexA(stop.FRT)mCD8.GFP (BL 57588), UAS-VP16::Hb/-;UAS-VP16::Hb (gift of C. Doe), UAS-svp1 1.12 (gift of M. Kowhi), Engrailed-GAL4 (BL 1973) (gift of M. Kowhi), UAS-hid,rpr (Zhou et al., 1997 (link)).

Meng J.L., Marshall Z.D., Lobb-Rabe M, & Heckscher E.S. (2019). How prolonged expression of Hunchback, a temporal transcription factor, re-wires locomotor circuits. eLife, 8, e46089.

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Drosophila Genetic Tools for Imaging

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The following Drosophila lines were used in this study: UAS-CD4-tdGFP (third chromosome insertion) (Han et al., 2011 (link)), UAS-nlsGFP (second chromosome insertion, Bloomington Stock Center), dpp-GAL4 (second chromosome insertion) (Roy et al., 2011 (link)), ppk-GAL4 (third chromosome insertion) (Kuo et al., 2005 (link)), and ppk-CD4-tdTomato (second chromosome insertion) (Han et al., 2011 (link)). w1118 was used as the wild-type strain. Unless noted otherwise, fly crosses were maintained at 25°C. The Drosophila transgenic of UAS-min-iSOG2 T2A H3.3-EGFP was generated in this study by using attP insertion on the second chromosome.

Makhijani K., To T.L., Ruiz-González R., Lafaye C., Royant A, & Shu X. (2017). Precision Optogenetic Tool for Selective Single- and Multiple-Cell Ablation in a Live Animal Model System. Cell chemical biology, 24(1), 110-119.

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Drosophila Genetics and Imaging Protocol

Cited 3 times

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All flies were raised at 25 °C with 12 h/12 h light/dark cycle unless noted. This study: htl-LexA, pyr-Gal4/CyO, htl>FRT>stop>FRT>Gal4, UAS-Pyr:GFP, UAS-Ths:GFP and LexO-Htl:mCherry. All new transgenic injections were performed by Rainbow Transgenic Flies, Inc. Bloomington Drosophila Stock Center: UAS-CD8:GFP, UAS-CD8:RFP, UAS-mCherryCAAX, LexO-CD2:GFP, UAS-Eb1:GFP, UAS-Lifeact:GFP, UAS-nls:GFP, UAS-nls:mCherry, htl-Gal4, ths-Gal4/CyO, UAS-Dia:GFP, UAS-ΔDAD-Dia:GFP, UAS-pyrRNAi, UAS-diaRNAi, hs-Flp, {nos-Cas9}ZH-2A, and w¹¹¹⁸. Vienna Drosophila Resource Center: htl:GFP^fTRG, UAS-htlRNAi, and UAS-thsRNAi. Other sources: LexO-nsyb:GFP^1–10, UAS-CD4:GFP¹¹^{20 (link)}. LexO-mCherryCAAX^{15 (link)}. dpp-Gal4/CyO, LexO-Fz:mCherry and 1151-Gal4 from Huang et al.^{36 (link)} (also see Supplementary Table 3).

Patel A., Wu Y., Han X., Su Y., Maugel T., Shroff H, & Roy S. (2022). Cytonemes coordinate asymmetric signaling and organization in the Drosophila muscle progenitor niche. Nature Communications, 13, 1185.

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Genetic Tools for Drosophila Muscle Development

Cited 1 time

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The sns and hbs double mutant sns^ZF1.4 hbs⁴⁵⁹ was generated for this work by recombining sns^ZF1.4 and hbs⁴⁵⁹, a loss-of-function allele of sns and hbs, respectively, onto the second chromosome. N^fa-g, UAS-Notch RNAi, Dl-lacZ, y w hsFLP, UAS-nlsGFP and Act5C>y+>Gal4 UAS-GFP were provided by the Bloomington Stock Center. rst-Gal4 was obtained from National Institute of Genetics Fly Stock Center (Japan). Other flies used: rst^F6-lacZ[31] (link), spa-Gal4[25] (link), UAS-N-cadherin[12] (link), sns^ZF1.4 and UAS-sns (gift of Susan Abmayr), UAS-N^ICD (gift of Cedric Wesley), UAS-N^ICD-lexA (gift of Toby Lieber), P[w+]36.1 and hbs⁴⁵⁹ (gift of Mary Baylies), UAS-hbs (gift of Helen Sink), GBE-Su(H)_m8-lacZ (N-lacZ) [22] (link), Gal-54[23] (link), UAS-rst (gift of Karl-F. Fischbach), UAS-kirre/duf (gift of Marc Ruiz-Gomez), UAS-Dl (gift of Marek Mlodzik) and hsFLP MKRS (gift of Matthew Freeman).

, & Bao S. (2014). Notch Controls Cell Adhesion in the Drosophila Eye. PLoS Genetics, 10(1), e1004087.

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Genetic Manipulation of Notch Signaling

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For genetic manipulations the Gal4 driver line 1151-Gal4 (Lingadahalli S. Shashidhara, Centre for Cellular and Molecular Biology, Hyderabad, India) was used and combined with UAS-N^ΔECD to provide constitutively active Notch (Fortini et al., 1993 (link), Rebay et al., 1993 (link)). These were combined with RNAi lines as listed in Table S4 and STAR Methods key resources table, including UAS-Hairless-RNAi (Bloomington Drosophila Stock Center, BL-27315), UAS-Mam-RNAi (BL 28046), UAS-Trr-RNAi (BL36916) or with UAS-MamDN to block Mam activity (Helms et al., 1999 (link)). Crosses were maintained at 25°C. Other lines used include NRE-GFP (Housden et al., 2012 (link)), DpE(spl)∂-8 (Chanet et al., 2009 (link)), UAS-nls-GFP (Bloomington 65402), Fkh::GFP (Bloomington 43951), SMRTR::YFP (DGRC 115513) (Lowe et al., 2014 (link)) and yw (Bloomington 1495). Further details are provided in Table S4.

Gomez-Lamarca M.J., Falo-Sanjuan J., Stojnic R., Abdul Rehman S., Muresan L., Jones M.L., Pillidge Z., Cerda-Moya G., Yuan Z., Baloul S., Valenti P., Bystricky K., Payre F., O'Holleran K., Kovall R, & Bray S.J. (2018). Activation of the Notch Signaling Pathway In Vivo Elicits Changes in CSL Nuclear Dynamics. Developmental Cell, 44(5), 611-623.e7.

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Drosophila Fly Line Acquisition for Research

Cited 1 time

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We ordered the following fly lines from the Bloomington Drosophila Stock Center (BDSC): C767-GAL4 (RRID:BDSC_30848), UAS-Dicer2 (RRID:BDSC_24650 and RRID:BDSC_24651), UAS-nlsGFP (RRID:BDSC_7032), UAS-mCD8::GFP (RRID:BDSC_5130), and DILP2-GAL4 (RRID:BDSC_37516). We ordered DH44-GAL4 (VT ID 039046) from the Vienna Drosophila Resource Center (VDRC) [25 ]. SIFa-GAL4 [8 (link)], kurs58-GAL4 (FBti0017957) [26 (link)] and Dilp2mCherry (FBti0202307) [5 (link)] were gifts from Amita Sehgal. C929-GAL4 (FBti0004282) [27 (link)] was a gift from Paul Taghert. We obtained RNAi lines for behavioral screening from the VDRC and the BDSC (see S1 File for a complete list of RNAi lines) [28 (link), 29 (link)].

Ruiz D., Bajwa S.T., Vanani N., Bajwa T.A, & Cavanaugh D.J. (2021). Slowpoke functions in circadian output cells to regulate rest:activity rhythms. PLoS ONE, 16(3), e0249215.

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