Genome-scale RNAi profiling of cell division in human tissue culture cells
Genome-scale RNAi profiling of cell division in human tissue culture cells
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DOI：
10.1038/ncb1659
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发表时间：
2007-12-01
影响因子：
21.3
通讯作者：
Buchholz, Frank
中科院分区：
文献类型：
--
作者：
Kittler, Ralf;Pelletier, Laurence;Buchholz, Frank
细胞分裂是所有生物体的基础。在这里，我们报告了在HeLa细胞中进行的基因组规模的RNA介导的干扰筛选，旨在识别对细胞分裂重要的人类基因。我们已经使用内切核酸酶准备的短干扰RNA文库来进行基因沉默，并使用DNA含量分析来识别导致细胞周期停滞或在沉默时改变倍性的基因。进行验证和二次分析，以产生每个基因的九个参数的功能丧失表型。这些表型特征允许通过结合层次聚类、跨物种分析和蛋白质组数据挖掘将基因分配到特定的功能类别。我们通过将新的功能归因于有丝分裂和胞质分裂中的基因来突出我们数据集的丰富性。特别是，我们确定了两个进化上保守的转录调控网络，它们管理着细胞质分裂。我们的工作提供了一个实验框架，可以从这个框架开始对人类细胞分裂所需的新基因进行系统分析。
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.