Uas lifeact gfp

Manufactured by Bloomington Drosophila Stock Center

Sourced in Australia, Germany, United States

UAS-Lifeact-GFP is a genetic construct that expresses the Lifeact peptide fused to the green fluorescent protein (GFP). Lifeact is a peptide that binds to actin filaments, allowing for visualization of the actin cytoskeleton within cells. When expressed, the UAS-Lifeact-GFP construct enables researchers to observe and study the dynamics of the actin cytoskeleton in Drosophila cells and tissues.

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UAS-GFP (63 citations)

8 protocols using uas lifeact gfp

Genetic Tools for Drosophila Cytoskeletal Analysis

Cited 4 times

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The following stocks were used: Oregon R, pyd^EX147 (Djiane et al., 2011 (link)), sns-GCN-nGFP (Zhuang et al., 2009 (link)), hand-GFP (Han et al., 2006 (link)), Fas3^MI03674 (GFP-tagged Fas3), Df(3L)vin3, CycA^C8LR1, UAS-Lifeact-GFP [Bloomington Drosophila Stock Center (BDSC), #59809, #6627, #2609 and #35544], CycA^C551 (this study, see below), sdk^CPTI000337, Rho1-GFP (Kyoto Drosophila Stock Center, #115107 and #110833), VT016847-Gal4 [Vienna Drosophila Resource Center (VDRC), #203048], UAS-PLCγ-PH-GFP (Pinal et al., 2006 (link)), UAS-Venus-Pbl (a gift from Stephen L. Gregory, University of Adelaide, Australia), UAS-sqh^DD (Mitonaka et al., 2007 (link)), Dia-GFP (Schmidt et al., 2021 (link)), sGMCA-GFP (Dutta et al., 2002 (link)), UAS-OCRL-GFP (Del Signore et al., 2017 (link)), UAS-aPKC^ΔN-GFP (Tian and Deng, 2008 (link)) and UAS-aPKC^CAAXDN (Sotillos et al., 2004 (link)).

Carrasco-Rando M., Culi J., Campuzano S, & Ruiz-Gómez M. (2023). An acytokinetic cell division creates PIP2-enriched membrane asymmetries leading to slit diaphragm assembly in Drosophila nephrocytes. Development (Cambridge, England), 150(18), dev201708.

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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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Drosophila Ovary Follicle Isolation

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Drosophila melanogaster were reared at 20–23 °C on standard medium with additional fresh yeast. 2–3 days old females were killed by crushing the head and thorax with tweezers without anaesthesia. The ovaries were dissected and single follicles of vitellogenic stages (S8–12) were isolated (see Fig. 1). The preparations were carried out in R-14 medium [57 (link)] which is best suited for in-vitro culture of Drosophila follicles [58 (link)].
In addition to wild-type (Oregon R), we used the Gal4/UAS system for the follicle-cell specific expression (Tj-Gal4; gift of S. Roth, Köln, Germany) of GFP-actin (UAS-Lifeact-GFP; Bloomington Stock Center, USA) and GFP-α-tubulin (UAS-αTub84B; Bloomington), respectively.

Weiß I, & Bohrmann J. (2019). Electrochemical gradients are involved in regulating cytoskeletal patterns during epithelial morphogenesis in the Drosophila ovary. BMC Developmental Biology, 19, 22.

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Drosophila Neuroblast Live Imaging

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Flies were raised in vials containing standard cornmeal-agar medium supplemented with baker’s yeast. Crosses were incubated at 25°C and larvae were grown for 4 days before being dissected for live imaging. Mutant chromosomes were balanced over Cyo:ActGFP or TM6B, Tb. The following mutant alleles and RNAi lines were used: Arp3^EP3640 (BL17149, Bloogminton),⁴² (link) Df(3L)Exel6112 (removes Arp3, BL7591, Bloogminton), SCAR^Δ37 FRT40A (BL8754, Bloomington),⁴⁰ (link) SCAR RNAi (BL36121, Bloomington). The following transgenes and fluorescent markers were used: Sqh:GFP,⁴³ (link) and UAS-cherry:Jupiter⁴⁴ (link) from C. Roubinet. UAS-PLCΔPH::GFP (BL39693, Bloomington), UAS-PH:mCherry (BL51658, Bloomington), UAS-LifeAct:GFP (BL58718, Bloomington), UAS-SCAR::GFP (from M. González-Gaitán). Transgenes were expressed using the neuroblast-specific driver worniu-Gal4.⁴⁵ (link)

Cazzagon G., Roubinet C, & Baum B. (2023). Polarized SCAR and the Arp2/3 complex regulate apical cortical remodeling in asymmetrically dividing neuroblasts. iScience, 26(7), 107129.

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Targeted Gene Expression in Drosophila

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Drosophila stocks were maintained at 25°C. The w¹¹¹⁸ strain was used as wild type. The targeted expression experiments were performed using the UAS-GAL4 system (Brand and Perrimon, 1993 (link)) on the following GAL4 and UAS lines: eme-GAL4 (kindly provided by R. Bodmer, Burnham Institute, San Diego, CA, USA); UAS-Rpr (BL5824), UAS-Antp (BL7301), UAS-Ubx (BL911), UAS-AbdA (BL912), UAS-AbdB (BL913), UAS-mcd8-GFP (BL32184) and UAS-LifeAct-GFP (BL58718) from Bloomington Stock Center; UAS-Tup RNAi (45859) and UAS-Eve RNAi (9284) from VDRC; UAS-Tin (kindly provided by M. Frasch, Erlangen-Nürnberg University, Germany); UAS-Lbe (Jagla et al., 1997 (link)); and UAS-Kr (a gift from G. Vorbrüggen, Max Planck Institute, Goettingen, Germany). The following additional lines were used: UAS-Antp RNAi (101774) and UAS-AbdA RNAi (106155) from VDRC; Antp25 (3020) from Bloomington Stock Center; and AbdAM1 (kindly provided by L. Perrin; Aix-Marseille University, France). The Tup-GFP line was kindly provided by Stephan Thor (Linköping University, Sweden), the Hand-GFP line was a gift from E. Olson (Utah University, Salt Lake City, USA) and the UAS-H2B::YFP line was kindly provided by Michel Gho (IBPS, Paris, France). UAS-Ubx was re-balanced with TM3, twi-lacZ, and homozygous mutant embryos were selected by absence of β-galactosidase staining.

Zmojdzian M., de Joussineau S., Da Ponte J.P, & Jagla K. (2018). Distinct subsets of Eve-positive pericardial cells stabilise cardiac outflow and contribute to Hox gene-triggered heart morphogenesis in Drosophila. Development (Cambridge, England), 145(2), dev158717.

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Drosophila Stem Cell Lineage Tracking

Cited 1 time

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We obtained hemolectinGal4; UAS-2xEGFP (BL30140), ubi-his2av::mRFP (BL23650), UAS-LifeActGFP (BL35544), and 10xUAS-IVS-myr-td::Eos (UAS-Eos) (BL32226) from the Bloomington Stock Center. esgGal4 (112304) was obtained from the Kyoto Drosophila Genomics and Genetics Resource (DGGR). The following stocks were gifts: mexGal4 (Carl Thummel), breathlessGal4, UAS-his2b::CFP (Yoshihiro Inoue), ubi-his2avD::YFP (Pavel Tomancak), and GBE-Su(H)-GFP::nls (Joaquin de Navascues), UAS-CD8-GFP, hs-flp¹²; tubGal4; FRT82, tubGal80 and FRT82 (David Bilder). The “fate sensor” line (esgGal4, UAS-his2b::CFP, GBE-Su(H)-GFP::nls; ubi-his2av::mRFP) was generated previously [12 (link)].

Koyama L.A., Aranda-Díaz A., Su Y.H., Balachandra S., Martin J.L., Ludington W.B., Huang K.C, & O’Brien L.E. (2020). Bellymount enables longitudinal, intravital imaging of abdominal organs and the gut microbiota in adult Drosophila. PLoS Biology, 18(1), e3000567.

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Genetic Tools for Drosophila Morphogenesis

Cited 1 time

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btl^LG19, bnl^P1, Df(2L)Exel6029, Df(3L)ri79c, Df(X)N73, da-GAL4, en-GAL4, UAS-myr-GFP, UAS-lifeact-GFP, AyGAL-FRT; UAS-lacZ, and hsFLP were obtained from Bloomington. btl-GAL4 was provided by S. Hayashi. UAS bnl^A1-1 and UAS bnl^A1-1 were provided by M. Krasnow. sage-GAL4 was created by our lab (Chung et al., 2009 (link)).

Cheng Y.L, & Andrew D.J. (2015). Extracellular Mipp1 activity confers migratory advantage to epithelial cells during collective migration. Cell reports, 13(10), 2174-2188.

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Drosophila Neurodegeneration Research Protocols

Cited 2 times

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Flies were maintained on cornmeal-molasses-yeast medium at 25 °C, 65% humidity, 12 hr light/dark cycle. The following strains were used in this study: UAS-Tau^WT and UAS-Tau^R406W obtained from Dr. Mel Feany (Ali et al., 2012 (link)); UAS-mitoGFP obtained from Dr. Hugo J. Bellen (Duncan Neurological Research Institute, Baylor College of Medicine); UAS-Nmnat-PD, UAS-Nmnat-PC, UAS-Nmnat-PD^WR generated from the lab (Ruan et al., 2015 (link); Zhai et al., 2006 (link)), GMR-GAL4, OK371-GAL4, and UAS-LifeAct-GFP obtained from Bloomington Stock Center.

Ma X., Zhu Y., Lu J., Xie J., Li C., Shin W.S., Qiang J., Liu J., Dou S., Xiao Y., Wang C., Jia C., Long H., Yang J., Fang Y., Jiang L., Zhang Y., Zhang S., Zhai R.G., Liu C, & Li D. (2020). Nicotinamide mononucleotide adenylyltransferase uses its NAD+ substrate-binding site to chaperone phosphorylated Tau. eLife, 9, e51859.

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