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

Ralf Kittler, Gabriele Putz, Laurence Pelletier, Ina Poser, Anne Kristin Heninger, David Drechsel, Steffi Fischer, Irena Konstantinova, Blanca Habermann, Hannes Grabner, Marie Laure Yaspo, Heinz Himmelbauer, Bernd Korn, Karla Neugenbaur, Maria Teresa Pisabarro, Frank Buchholz

Research output: Contribution to journalArticle

321 Scopus citations

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 molecules. In invertebrates, long dsRNAs have been used for genome-wide screens and have provided insights into gene functions. 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 silencing. Here we report the generation of a genome-scale library of endoribonuclease-prepared short interfering (esi)RNAs 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.

Original languageEnglish (US)
Pages (from-to)1036-1040
Number of pages5
JournalNature
Volume432
Issue number7020
DOIs
StatePublished - Dec 23 2004

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    Kittler, R., Putz, G., Pelletier, L., Poser, I., Heninger, A. K., Drechsel, D., Fischer, S., Konstantinova, I., Habermann, B., Grabner, H., Yaspo, M. L., Himmelbauer, H., Korn, B., Neugenbaur, K., Pisabarro, M. T., & Buchholz, F. (2004). An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division. Nature, 432(7020), 1036-1040. https://doi.org/10.1038/nature03159