GenomeRNAi - a database for RNAi phenotypes and reagents
TitleScreen TitleAssayBiomodelSpecies
An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division. Kittler et al. (2004) Cell cycle Cell viability / cell cycleHeLa (cervix carcinom)H. sapiens

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.

Screen details


Stable Id: GR00097-A-0
Screen title: Cell cycle
Assay: Cell viability / cell cycle
Method: High content (microscopy)
Scope:
Screen type: Cell-based
Species: Homo sapiens
Biosource: Cell line
Biomodel: HeLa (cervix carcinom)
Library: , esiRNA (Kittler et al.)
Reagent type: esiRNA
Score type: Z-score
Cutoff: >= 1.645 / <= 1.645
Notes: