1. Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany
2. Scionics Computer Innovation, GmbH, Pfotenhauerstrasse 110, D-01307 Dresden, Germany
3. Max Planck Institute for Molecular Genetics, Ihnestrasse 73, D-14195 Berlin-Dahlem, Germany
4. RZPD-Ressourcenzentrum für Genomforschung, Im Neuenheimer Feld 506, D-69120 Heidelberg, Germany
5. Present address: TU Dresden, Biotechnologisches Zentrum, Tatzberg 47-51, D-01307 Dresden, Germany

Correspondence to: Frank Buchholz¹ Email: buchholz@mpi-cbg.de
The sequences of KIAA1387 are deposited in GenBank under accession numbers AY825268 and AY825269.

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^{1, 2, 3, 4}. In invertebrates, long dsRNAs have been used for genome-wide screens and have provided insights into gene functions^{5, 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 silencing^{9, 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.

MORE ARTICLES LIKE THIS

These links to content published by NPG are automatically generated.