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

Cell division is a fundamental process for multicellula organisation. Here we report a genome-scale RNAi screen in HeLa cells designed to identify human genes important for cell division. We utilized a library of endoribonuclease-prepared siRNAs (esiRNAs) for gene-silencing and used DNA content analysis to uncover genes that induced cell cycle arrest or altered ploidy upon knockdown. Rigorous validation and secondary assays were preformed to generate a nine-parameter fingerprint 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 describing novel functions to genes in mitosis and cytokinesis. In particular, we identified two evolutionarily conserved transcriptional regulatory networks that govern cytokinesis in mammalian cells. Our work provides an experimental framework from which the systematic analysis of novel genes necessary for cell division in human cells can begin. Keywords: time course An esiRNA targeting LIN54 was transfected into HCT116 cells. RNA was harvested at 24, 48, and 72 hours post-transfection for microarray analysis. Transcripts regulated at least 1.5-fold, and with a p-value<0.01 were identified as signature transcripts.