A gene-specific T2A-GAL4 library for Drosophila

  • Pei-Tseng Lee
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Jonathan Zirin
    Department of Genetics, Harvard Medical School, Boston, United States
  • Oguz Kanca
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Wen-Wen Lin
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Karen L Schulze
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • David Li-Kroeger
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Rong Tao
    Department of Genetics, Harvard Medical School, Boston, United States
  • Colby Devereaux
    Department of Genetics, Harvard Medical School, Boston, United States
  • Yanhui Hu
    Department of Genetics, Harvard Medical School, Boston, United States
  • Verena Chung
    Department of Genetics, Harvard Medical School, Boston, United States
  • Ying Fang
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Yuchun He
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Hongling Pan
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Ming Ge
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Zhongyuan Zuo
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Benjamin E Housden
    Department of Genetics, Harvard Medical School, Boston, United States
  • Stephanie E Mohr
    Department of Genetics, Harvard Medical School, Boston, United States
  • Shinya Yamamoto
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
  • Robert W Levis
    Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
  • Allan C Spradling
    Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
  • Norbert Perrimon
    Department of Genetics, Harvard Medical School, Boston, United States
  • Hugo J Bellen
    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States

説明

<jats:p>We generated a library of ~1000 Drosophila stocks in which we inserted a construct in the intron of genes allowing expression of GAL4 under control of endogenous promoters while arresting transcription with a polyadenylation signal 3’ of the GAL4. This allows numerous applications. First, ~90% of insertions in essential genes cause a severe loss-of-function phenotype, an effective way to mutagenize genes. Interestingly, 12/14 chromosomes engineered through CRISPR do not carry second-site lethal mutations. Second, 26/36 (70%) of lethal insertions tested are rescued with a single UAS-cDNA construct. Third, loss-of-function phenotypes associated with many GAL4 insertions can be reverted by excision with UAS-flippase. Fourth, GAL4 driven UAS-GFP/RFP reports tissue and cell-type specificity of gene expression with high sensitivity. We report the expression of hundreds of genes not previously reported. Finally, inserted cassettes can be replaced with GFP or any DNA. These stocks comprise a powerful resource for assessing gene function.</jats:p>

収録刊行物

  • eLife

    eLife 7 e35574-, 2018-03-22

    eLife Sciences Publications, Ltd

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