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An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division

Author

Listed:
  • Ralf Kittler

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Gabriele Putz

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Laurence Pelletier

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Ina Poser

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Anne-Kristin Heninger

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • David Drechsel

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Steffi Fischer

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Irena Konstantinova

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Bianca Habermann

    (Scionics Computer Innovation, GmbH)

  • Hannes Grabner

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Marie-Laure Yaspo

    (Max Planck Institute for Molecular Genetics)

  • Heinz Himmelbauer

    (Max Planck Institute for Molecular Genetics)

  • Bernd Korn

    (RZPD-Ressourcenzentrum für Genomforschung)

  • Karla Neugebauer

    (Max Planck Institute for Molecular Cell Biology and Genetics)

  • Maria Teresa Pisabarro

    (Max Planck Institute for Molecular Cell Biology and Genetics
    Biotechnologisches Zentrum)

  • Frank Buchholz

    (Max Planck Institute for Molecular Cell Biology and Genetics)

Abstract

RNA interference (RNAi) is an evolutionarily conserved defence mechanism whereby genes are specifically silenced through degradation of messenger RNAs; this process is mediated by homologous double-stranded (ds)RNA molecules1,2,3,4. In invertebrates, long dsRNAs have been used for genome-wide screens and have provided insights into gene functions5,6,7,8. Because long dsRNA triggers a nonspecific interferon response in many vertebrates, short interfering (si)RNA or short hairpin (sh)RNAs must be used for these organisms to ensure specific gene silencing9,10,11. Here we report the generation of a genome-scale library of endoribonuclease-prepared short interfering (esi)RNAs12 from a sequence-verified complementary DNA collection representing 15,497 human genes. We used 5,305 esiRNAs from this library to screen for genes required for cell division in HeLa cells. Using a primary high-throughput cell viability screen followed by a secondary high content videomicroscopy assay, we identified 37 genes required for cell division. These include several splicing factors for which knockdown generates mitotic spindle defects. In addition, a putative nuclear-export terminator was found to speed up cell proliferation and mitotic progression after knockdown. Thus, our study uncovers new aspects of cell division and establishes esiRNA as a versatile approach for genomic RNAi screens in mammalian cells.

Suggested Citation

  • Ralf Kittler & Gabriele Putz & Laurence Pelletier & Ina Poser & Anne-Kristin Heninger & David Drechsel & Steffi Fischer & Irena Konstantinova & Bianca Habermann & Hannes Grabner & Marie-Laure Yaspo & , 2004. "An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division," Nature, Nature, vol. 432(7020), pages 1036-1040, December.
    • Handle: RePEc:nat:nature:v:432:y:2004:i:7020:d:10.1038_nature03159
    DOI: 10.1038/nature03159
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    Cited by:

    1. Victoria O. Shender & Ksenia S. Anufrieva & Polina V. Shnaider & Georgij P. Arapidi & Marat S. Pavlyukov & Olga M. Ivanova & Irina K. Malyants & Grigory A. Stepanov & Evgenii Zhuravlev & Rustam H. Zig, 2024. "Therapy-induced secretion of spliceosomal components mediates pro-survival crosstalk between ovarian cancer cells," Nature Communications, Nature, vol. 15(1), pages 1-26, December.

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