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

Nature. 2004 Dec 23;432(7020):1036-40. doi: 10.1038/nature03159.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Division / genetics*
  • Cell Proliferation
  • Cell Survival
  • Cytokinesis / genetics
  • Endoribonucleases / metabolism*
  • Gene Library*
  • Genes, Essential / genetics*
  • Genomics / methods*
  • HeLa Cells
  • Humans
  • Microscopy, Video
  • Mitosis / genetics
  • Molecular Sequence Data
  • Phenotype
  • RNA Interference*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism*
  • Spindle Apparatus / physiology


  • RNA, Small Interfering
  • Endoribonucleases

Associated data

  • GENBANK/AY825268
  • GENBANK/AY825269
  • RefSeq/NM_004048