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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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