Article abstract

Nature Cell Biology 9, 1401 - 1412 (2007)
Published online: 11 November 2007 | doi:10.1038/ncb1659

Genome-scale RNAi profiling of cell division in human tissue culture cells

Ralf Kittler1,6, Laurence Pelletier1,7, Anne-Kristine Heninger1, Mikolaj Slabicki1, Mirko Theis1, Lukasz Miroslaw1, Ina Poser1, Steffen Lawo7, Hannes Grabner1,2, Karol Kozak1,2, Jan Wagner1,2, Vineeth Surendranath1, Constance Richter1, Wayne Bowen3, Aimee L. Jackson4, Bianca Habermann1,5, Anthony A. Hyman1 & Frank Buchholz1

Cell division is fundamental for all organisms. Here we report a genome-scale RNA-mediated interference screen in HeLa cells designed to identify human genes that are important for cell division. We have used a library of endoribonuclease-prepared short interfering RNAs for gene silencing and have used DNA content analysis to identify genes that induced cell cycle arrest or altered ploidy on silencing. Validation and secondary assays were performed to generate a nine-parameter loss-of-function phenoprint for each of the genes. These phenotypic signatures allowed the assignment of genes to specific functional classes by combining hierarchical clustering, cross-species analysis and proteomic data mining. We highlight the richness of our dataset by ascribing novel functions to genes in mitosis and cytokinesis. In particular, we identify two evolutionarily conserved transcriptional regulatory networks that govern cytokinesis. Our work provides an experimental framework from which the systematic analysis of novel genes necessary for cell division in human cells can begin.

  1. Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany.
  2. Technology Development Studio, Pfotenhauerstrasse 108, D-01307 Dresden, Germany.
  3. TTP LabTech Ltd, Melbourn Science Park, Melbourn, Hertfordshire SG8 6EE, UK.
  4. Rosetta Inpharmatics LLC, 401 Terry Avenue N, Seattle, Washington 98109, USA.
  5. Scionics Computer Innovation GmbH, Pfotenhauerstrasse 110, D-01307 Dresden, Germany.
  6. Present address: Department of Human Genetics, University of Chicago, CLSB, 920 E. 58th Street, Chicago, Illinois 60637, USA.
  7. Present address: Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario M5G 1X5, Canada.

Correspondence to: Frank Buchholz1 e-mail:


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