An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
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.
Details
Original language | English |
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Pages (from-to) | 1036-1040 |
Number of pages | 5 |
Journal | Nature |
Volume | 432 |
Issue number | 7020 |
Publication status | Published - 23 Dec 2004 |
Peer-reviewed | Yes |
External IDs
Scopus | 19944367565 |
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Keywords
Keywords
- 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, Spindle Apparatus/physiology