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          Institute: MPI für molekulare Genetik     Collection: Department of Vertebrate Genomics     Display Documents



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ID: 230601.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division
Authors:Kittler, Ralf; Putz, Gabriele; Pelletier, Laurence; Poser, Ina; Heninger, Anne-Kristin; Drechsel, David; Fischer, Steffi; Konstantinova, Irena; Habermann, Bianca; Grabner, Hannes; Yaspo, Marie-Laure; Himmelbauer, Heinz; Korn, Bernd; Neugebauer, Karla; Pisabarro, Maria Teresa; Buchholz, Frank
Language:English
Date of Publication (YYYY-MM-DD):2004-12-23
Title of Journal:Nature
Volume:432
Issue / Number:7020
Start Page:1036
End Page:1040
Copyright:© 2004 Nature Publishing Group
Review Status:not specified
Audience:Experts Only
Abstract / Description: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.
External Publication Status:published
Document Type:Article
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany;
Scionics Computer Innovation, GmbH, Pfotenhauerstrasse 110, D-01307 Dresden, Germany;
RZPD-Ressourcenzentrum für Genomforschung, Im Neuenheimer Feld 506, D-69120 Heidelberg, Germany
Identifiers:ISSN:0028-0836
DOI:10.1038/nature03159
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