Kittler Ralf, Pelletier Laurence, Heninger Anne-Kristine, Slabicki Mikolaj, Theis Mirko, Miroslaw Lukasz, Poser Ina, Lawo Steffen, Grabner Hannes, Kozak Karol, Wagner Jan, Surendranath Vineeth, Richter Constance, Bowen Wayne, Jackson Aimee L, Habermann Bianca, Hyman Anthony A, Buchholz Frank
Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany.
Nat Cell Biol. 2007 Dec;9(12):1401-12. doi: 10.1038/ncb1659. Epub 2007 Nov 11.
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.
细胞分裂是所有生物体的基础。在此，我们报告了一项在HeLa细胞中进行的全基因组规模的RNA介导的干扰筛选，旨在鉴定对细胞分裂重要的人类基因。我们使用了一个由核糖核酸酶制备的短发夹RNA文库进行基因沉默，并使用DNA含量分析来鉴定在沉默时诱导细胞周期停滞或改变倍性的基因。进行了验证和二次分析，以生成每个基因的九参数功能丧失表型印记。通过结合层次聚类、跨物种分析和蛋白质组数据挖掘，这些表型特征允许将基因分配到特定的功能类别。我们通过赋予有丝分裂和胞质分裂中基因的新功能来突出我们数据集的丰富性。特别是，我们鉴定出两个控制胞质分裂的进化保守转录调控网络。我们的工作提供了一个实验框架，从该框架可以开始对人类细胞中细胞分裂所需的新基因进行系统分析。
