Research
Whole genome functional analysis identifies novel components required for mitotic spindle integrity in human cells
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* Corresponding authors: Sumit K Chanda schanda@burnham.org - Jeremy S Caldwell jcaldwell@gnf.org
1 Genomics Institute of Novartis Research Foundation, John Jay Hopkins Drive, San Diego, California 92121, USA
2 Department of Physiology and Biophysics, Albert Einstein College of Medicine, 1300 Marris Park Avenue, Bronx, New York 10461, USA
3 Novartis Institute for Biomedical Research Inc., Mass Avenue, Cambridge, Massachusetts 02139, USA
4 Novartis Pharma AG, Postfach, CH-4002 Basel, Switzerland
5 Infectious & Inflammatory Disease Center, Burnham Institute for Medical Research, North Torrey Pines Road, La Jolla, California 92037, USA
Genome Biology 2008, 9:R44 doi:10.1186/gb-2008-9-2-r44
Published: 26 February 2008Abstract
Background
The mitotic spindle is a complex mechanical apparatus required for accurate segregation of sister chromosomes during mitosis. We designed a genetic screen using automated microscopy to discover factors essential for mitotic progression. Using a RNA interference library of 49,164 double-stranded RNAs targeting 23,835 human genes, we performed a loss of function screen to look for small interfering RNAs that arrest cells in metaphase.
Results
Here we report the identification of genes that, when suppressed, result in structural defects in the mitotic spindle leading to bent, twisted, monopolar, or multipolar spindles, and cause cell cycle arrest. We further describe a novel analysis methodology for large-scale RNA interference datasets that relies on supervised clustering of these genes based on Gene Ontology, protein families, tissue expression, and protein-protein interactions.
Conclusion
This approach was utilized to classify functionally the identified genes in discrete mitotic processes. We confirmed the identity for a subset of these genes and examined more closely their mechanical role in spindle architecture.