Uas kir2

Manufactured by Bloomington Drosophila Stock Center

Sourced in United States

The UAS-Kir2.1 is a genetic tool used in Drosophila research. It is a transgenic construct that expresses the inwardly rectifying potassium channel Kir2.1 under the control of the UAS promoter. This allows for the spatially and temporally controlled expression of Kir2.1 in specific cell types or tissues, which can be used to study their physiological functions.

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8 protocols using uas kir2

Genetic Tools for Drosophila Gustatory Receptors

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Flies were raised on standard cornmeal fly food at 25°C and 70% relative humidity. The following fly lines were used: Gr5a-GAL4, Gr43a-GAL4, Gr64a-GAL4, Gr61a-GAL4, Gr64c-GAL4, Gr64d-GAL4, and Gr64e-GAL4 (ref^{16 (link)}); Gr64f^LexA and Gr43a^GAL4 (ref^{13 (link)}); UAS-GCaMP3 (ref⁴³); UAS-KIR2.1 (ref^{37 (link)}); UAS-TdTomato and UAS-GFP (Bloomington stock center); poxn^Δ^M22-B5 (ref^{44 (link)}); poxn⁷⁰ (ref ^{28 (link)}).

LeDue E.E., Chen Y.C., Jung A.Y., Dahanukar A, & Gordon M.D. (2015). Pharyngeal sense organs drive robust sugar consumption in Drosophila: Pharyngeal taste in adult Drosophila. Nature communications, 6, 6667.

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Investigating Drosophila Circadian Rhythms

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Flies were raised in a 12 h:12 h light:dark (LD) cycle at 25°C in vials containing standard cornmeal medium. For these experiments, we use the following stocks: w¹¹¹⁸ (RRID:BDSC_5905), UAS-Kir2.1 (Nitabach et al., 2002 (link)), UAS-dTrpA1 (Rosenzweig et al., 2008 (link)), UAS-mCD8GFP, Clk856-GAL4 (Gummadova et al., 2009 (link)) and OK107-GAL4 (MB) driver) that were obtained from the Bloomington Stock Center. We used the same group of enhancer trap lines used by Gorostiza et al. (2014) (link): 3-86-GAL4, 11-8-GAL4, 4-12-GAL4, 4-93-GAL4, 5-133-GAL4, 4-59-GAL4, 5-43-GAL4 and 7-49-GAL4. These lines were a gift from U. Heberlein (Janelia Farm, USA). For the optical imaging experiments we used the following fly lines: pdf-LexA (Shang et al., 2008 (link)), UAS-GCaMP3 (Tian et al., 2009 (link)), LexAop-P2X2, pdf-GAL4 (Renn et al., 1999 (link)) (RRID:BDSC_6900), Clk4.1-GAL4 (Zhang et al., 2010 (link)), Clk4.5-GAL4, Mai179-GAL4>pdf-GAL80 and tim-GAL4>pdf-GAL80 (Emery et al., 1998 (link)) (RRID:BDSC_7126). We generated the experimental fly lines crossing the different GAL4s to the pdf-LexA,UAS-GCaMP3>LexAop-P2X2 line. The UAS-GCaMP3 was obtained from Janelia Farm and the LexAop-P2X2 was a gift from O. Shafer (University of Michigan) (Yao et al., 2012 (link)). All experimental protocols were performed in accordance with relevant guidelines and ethical regulations of our institution.

Pírez N., Bernabei-Cornejo S.G., Fernandez-Acosta M., Duhart J.M, & Ceriani M.F. (2018). Contribution of non-circadian neurons to the temporal organization of locomotor activity. Biology Open, 8(1), bio039628.

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Drosophila Rearing and Genetic Toolkit

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Flies were reared on standard cornmeal-yeast medium under a 12 hr:12 hr dark:light cycle at 25°C and 60% humidity. Flies carrying a dTrpA1 transgene were raised at 21°C. UAS-TNTE and UAS-impTNT were kindly provided by Dr Cahir O’Kane (University of Cambridge). UAS-dTRPA1 was a gift from Dr Paul Garrity (Brandeis University). Dilp2-GAL4 line, trans-Tango line, and elav-GS line were provided by Dr Yi Zhong (Tsinghua University), DskFlp and DskRNAi lines were provided by Dr Yufeng Pan (Southeast University). UAS > stop > myr::GFP (pJFRC41 in attP5) was a gift from Gerald Rubin, UAS > stop > kir^eGFP was provided by Dr Yi Rao, pC1-ss1 and pC1-ss2 were provided by Dr Kaiyu Wang. The following lines were obtained from the Bloomington Drosophila Stock Center: R71G01-GAL4 (BL#39599), R71G01-LexA (BL#54733), TβH-GAL4 (BL#45904), TRIC line (BL#61679), UAS-Kir2.1 (BL#6595 and BL#6596), UAS-mCD8::GFP (BL#5137), UAS > stop > dTrpA^myrc (BL#66871). Lexo-CD4-spGFP11/CyO; UAS-CD4-spGFP1-10/Tb was previously described (Gordon and Scott, 2009 (link)).

Wang T., Jing B., Deng B., Shi K., Li J., Ma B., Wu F, & Zhou C. (2022). Drosulfakinin signaling modulates female sexual receptivity in Drosophila. eLife, 11, e76025.

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Genetic Tools for Drosophila Gustatory Receptors

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Drosophila Genetic Tools and Manipulations

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Animals were raised on standard medium as used by the Bloomington Drosophila Stock Center. All animals were raised at 25°C, except ap>rpk‐RNAi crosses, which were raised at 18°C to reduce lethality. Crosses using GAL80^TS and the hh^TS2 allele were raised at 18°C and then upshifted to 30°C at the reported timepoints.
Stocks used in this study include the following: hsFLP;; act<stop<Gal4, UAS‐RFP/S‐T, yw;ap‐Gal4/Cy0; TM2/TM6B,;;ap‐Gal4,;ptc>RFP;, w;;UAS‐Ci3 M,;; 71B‐Gal4, and w;UAS‐Kir2.1; UAS‐Kir2.1/TM6B (from Kristin Scott, UC Berkeley, USA).
Stocks obtained from the Bloomington Stock Center (Bloomington, IN, USA) include UAS‐ArcLight (BL:51056), UAS‐rpk‐RNAi; (BL:39053, 25847—data using BL:39053 are shown and confirmed with BL:25847), UAS‐NaChBac; (BL:9466), UAS‐ChR2::mCherry; (BL:28995), UAS‐ReaChR::Citrine; (BL:53741), UAS‐ChloC::tdTom (BL:76328), en‐Gal4; (BL:6356),; UAS‐Ptc‐RNAi; (BL:55686).

Emmons‐Bell M, & Hariharan I.K. (2021). Membrane potential regulates Hedgehog signalling in the Drosophila wing imaginal disc. EMBO Reports, 22(4), e51861.

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Drosophila Husbandry and Genotypes

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Flies were raised on standard cornmeal-dextrose diet at 25°C and 50% relative humidity in a 12:12 light:dark cycle. For the diet, yellow cornmeal (57020, Quaker), dextrose (G8270, Sigma), inactive dry yeast (75570, Lynside), propionic acid (402907, Sigma) and tegosept (20–258, Apex chemical and reagent) were used. D. melanogaster flies were red-eyed Canton-S. Other D. melanogaster genotypes were obtained from the Bloomington Drosophila Stock Center: UAS-Kir2.1 (BL91802), Gr64f-Gal4 (BL27883), Gr32a-Gal4 (BL57622), Gr33a¹ (BL31427), Ir56d¹ (BL81249), Ir76b¹ (BL51309), orco¹ (BL23129), vas-Cas9 (BL# 51324), w¹¹¹⁸ (BL5905). wCS is w¹¹¹⁸ backcrossed to Canton-S. D. simulans (w[501] 14021–0251.195) and D. sechellia (14021–0248.25) were obtained from the Drosophila Species Stock Center. ΔGr64a-f flies were generously shared by Dr. Seok Jun Moon. Ir47a¹ mutants were generated in the laboratory for this study.

Pitzurra L., Cherniak R., Giammarioli M., Perito S., Bistoni F, & Vecchiarelli A. (2000). Early Induction of Interleukin-12 by Human Monocytes Exposed to Cryptococcus neoformans Mannoproteins. Infection and Immunity, 68(2), 558-563.

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Fly Genetic Tools for Sensory Neuron Analysis

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Or42b-Gal4, Or42b-Gal80, Nan-Gal4, Ir31a-Gal4, Ir41a-Gal4, Ir40a-Gal4, Ir75a-Gal4, Ir8a-Gal4, R11F02-Gal4, y¹,w^*,UAS-mCD8-RFP,LexAop2-mCD8-GFP, Orco², Or42b^EY14886, piezo^KO, nompC¹, IR68a^MB05565, IR25a², Df(2R)7094, LexAop-GCaMP6m, LexAop-Gal80, UAS-Kir2.1, Orco-RFP, UAS-RedStinger, UAS-CD4-tdGFP, UAS-GCaMP6m and UAS-tdTomato were obtained from the Bloomington Stock Center. UAS-Tmem63-RNAi and UAS-Dicer2 flies were obtained from the Vienna Drosophila Resource Center. UAS-ReaperHid was a gift from Hermann Steller lab. Ir68a-Gal4 was a gift from Paul Garrity lab. iav¹, tmc^GAL4, nompC^f00914 and UAS-pzl-RNAi were gifts from Wei Zhang lab. UAS-ppk-RNAi and UAS-nan-RNAi were from Tsinghua Fly Center. ppk28^Δ was from the Core Facility of Drosophila Resource and Technology in Shanghai Institutes of Biochemistry and Cell Biology. The flies were raised on standard medium at 25 °C and 60% humidity under a 12 h/ 12 h light-dark cycle.

Li S., Li B., Gao L., Wang J, & Yan Z. (2022). Humidity response in Drosophila olfactory sensory neurons requires the mechanosensitive channel TMEM63. Nature Communications, 13, 3814.

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Drosophila Neurophysiology Experimental Protocol

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Flies were maintained on a media of sucrose, yeast, molasses, and agar under 12:12 LD cycles at 25 °C. One- to 3-day-old female flies were separated and maintained on standard cornmeal-yeast medium under 12:12 LD cycles at 25 °C for 4 nights before experiments began. Clk[out] (56754), per^s (80919), pdf-Gal4 (6899), pBDP (pBDP-Gal4Uw)(68384), pBDP split (p65-AD Uw; Gal4-DBD Uw) (79603), R23E10-Gal4 (49032), R69F08-Gal4 (39499), R58H05 p59AD (70750), R48H04 DBD (69353) pdf-Gal80 (80940), R51H05 p65AD (70720), R18H11 DBD (69017), R92H07-Gal4 (40633), R52B02-Gal4 (38814), R20D01-Gal4 (48889), BRPstar (55751),UAS-GCaMP6s (42746), UAS-TNT (28838), UAS-kir2.1 (6596) and UAS-hid (65403) were obtained from the Bloomington Drosophila Stock Center. mGluR-RNAi (1793) was obtained from Vienna Drosophila Resource Center. MB122B and 20xUas-IVS-Syn-GFP was obtained from Janelia Farm.

Andreani T., Rosensweig C., Sisobhan S., Ogunlana E., Kath W, & Allada R. (2022). Circadian programming of the ellipsoid body sleep homeostat in Drosophila. eLife, 11, e74327.

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