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A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila

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  • Georg Dietzl

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria
    Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

  • Doris Chen

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria)

  • Frank Schnorrer

    (Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

  • Kuan-Chung Su

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria)

  • Yulia Barinova

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria)

  • Michaela Fellner

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria
    Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

  • Beate Gasser

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria)

  • Kaolin Kinsey

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria
    Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

  • Silvia Oppel

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria
    Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

  • Susanne Scheiblauer

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria)

  • Africa Couto

    (Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

  • Vincent Marra

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria)

  • Krystyna Keleman

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria
    Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

  • Barry J. Dickson

    (Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr. Bohr-Gasse 3-5, A-1030 Vienna, Austria
    Research Institute of Molecular Pathology (IMP) Dr. Bohr-Gasse 7, A-1030 Vienna, Austria)

Abstract

Forward genetic screens in model organisms have provided important insights into numerous aspects of development, physiology and pathology. With the availability of complete genome sequences and the introduction of RNA-mediated gene interference (RNAi), systematic reverse genetic screens are now also possible. Until now, such genome-wide RNAi screens have mostly been restricted to cultured cells and ubiquitous gene inactivation in Caenorhabditis elegans. This powerful approach has not yet been applied in a tissue-specific manner. Here we report the generation and validation of a genome-wide library of Drosophila melanogaster RNAi transgenes, enabling the conditional inactivation of gene function in specific tissues of the intact organism. Our RNAi transgenes consist of short gene fragments cloned as inverted repeats and expressed using the binary GAL4/UAS system. We generated 22,270 transgenic lines, covering 88% of the predicted protein-coding genes in the Drosophila genome. Molecular and phenotypic assays indicate that the majority of these transgenes are functional. Our transgenic RNAi library thus opens up the prospect of systematically analysing gene functions in any tissue and at any stage of the Drosophila lifespan.

Suggested Citation

  • Georg Dietzl & Doris Chen & Frank Schnorrer & Kuan-Chung Su & Yulia Barinova & Michaela Fellner & Beate Gasser & Kaolin Kinsey & Silvia Oppel & Susanne Scheiblauer & Africa Couto & Vincent Marra & Kry, 2007. "A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila," Nature, Nature, vol. 448(7150), pages 151-156, July.
  • Handle: RePEc:nat:nature:v:448:y:2007:i:7150:d:10.1038_nature05954
    DOI: 10.1038/nature05954
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    Cited by:

    1. Aviel Even & Giovanni Morelli & Silvia Turchetto & Michal Shilian & Romain Le Bail & Sophie Laguesse & Nathalie Krusy & Ariel Brisker & Alexander Brandis & Shani Inbar & Alain Chariot & Frédéric Saudo, 2021. "ATP-citrate lyase promotes axonal transport across species," Nature Communications, Nature, vol. 12(1), pages 1-14, December.
    2. Olga Kubrak & Anne F. Jørgensen & Takashi Koyama & Mette Lassen & Stanislav Nagy & Jacob Hald & Gianluca Mazzoni & Dennis Madsen & Jacob B. Hansen & Martin Røssel Larsen & Michael J. Texada & Jakob L., 2024. "LGR signaling mediates muscle-adipose tissue crosstalk and protects against diet-induced insulin resistance," Nature Communications, Nature, vol. 15(1), pages 1-22, December.
    3. Olga Kubrak & Takashi Koyama & Nadja Ahrentløv & Line Jensen & Alina Malita & Muhammad T. Naseem & Mette Lassen & Stanislav Nagy & Michael J. Texada & Kenneth V. Halberg & Kim Rewitz, 2022. "The gut hormone Allatostatin C/Somatostatin regulates food intake and metabolic homeostasis under nutrient stress," Nature Communications, Nature, vol. 13(1), pages 1-17, December.
    4. Hiroyuki Uechi & Kazuki Fukushima & Ryota Shirasawa & Sayaka Sekine & Erina Kuranaga, 2022. "Inhibition of negative feedback for persistent epithelial cell–cell junction contraction by p21-activated kinase 3," Nature Communications, Nature, vol. 13(1), pages 1-12, December.
    5. Michaela Joyce & Federica A. Falconio & Laurence Blackhurst & Lucia Prieto-Godino & Alice S. French & Giorgio F. Gilestro, 2024. "Divergent evolution of sleep in Drosophila species," Nature Communications, Nature, vol. 15(1), pages 1-11, December.
    6. Eyal Rozenfeld & Nadine Ehmann & Julia E. Manoim & Robert J. Kittel & Moshe Parnas, 2023. "Homeostatic synaptic plasticity rescues neural coding reliability," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    7. Zachary T. Spencer & Victoria H. Ng & Hassina Benchabane & Ghalia Saad Siddiqui & Deepesh Duwadi & Ben Maines & Jamal M. Bryant & Anna Schwarzkopf & Kai Yuan & Sara N. Kassel & Anant Mishra & Ashley P, 2023. "The USP46 deubiquitylase complex increases Wingless/Wnt signaling strength by stabilizing Arrow/LRP6," Nature Communications, Nature, vol. 14(1), pages 1-17, December.

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