Cancer proliferation gene discovery through functional Genomics
Cancer proliferation gene discovery through functional Genomics
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DOI：
10.1126/science.1149200
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发表时间：
2008-02-01
期刊：
影响因子：
56.9
通讯作者：
Elledge, Stephen J.
中科院分区：
文献类型：
--
作者：
Schlabach, Michael R.;Luo, Ji;Elledge, Stephen J.
逆转录病毒短发夹RNA（shRNA）介导的哺乳动物细胞基因筛选是发现功能缺失表型的有力工具。我们描述了一种高度并行的多重方法，用于使用半发夹条形码进行微阵列解卷积来筛选大量的shRNA池。我们针对影响癌细胞和正常细胞增殖及活力的shRNA进行了缺失筛选。我们发现许多针对所有受检细胞的核心细胞过程（如细胞周期和蛋白质翻译）的shRNA具有抗增殖作用。此外，我们还鉴定了在不同细胞系中增殖和存活选择性所需的基因。我们的平台能够快速且经济高效地进行全基因组筛选，以鉴定用于靶点发现的癌症增殖和存活基因。此类工作是对癌症基因组图谱的补充，并提供了癌细胞的另一种功能性视角。
Retroviral short hairpin RNA (shRNA) - mediated genetic screens in mammalian cells are powerful tools for discovering loss- of- function phenotypes. We describe a highly parallel multiplex methodology for screening large pools of shRNAs using half- hairpin barcodes for microarray deconvolution. We carried out dropout screens for shRNAs that affect cell proliferation and viability in cancer cells and normal cells. We identified many shRNAs to be antiproliferative that target core cellular processes, such as the cell cycle and protein translation, in all cells examined. Moreover, we identified genes that are selectively required for proliferation and survival in different cell lines. Our platform enables rapid and cost- effective genome-wide screens to identify cancer proliferation and survival genes for target discovery. Such efforts are complementary to the Cancer Genome Atlas and provide an alternative functional view of cancer cells.