Wed, 20 Aug 2008 23:25:31 BST CiteULike: jyuh's Grenier http://www.citeulike.org/user/jyuh/author/Grenier CiteULike.org en-gb Copyright © 2004-2008 citeulike.org http://www.citeulike.org/user/jyuh/article/1616351 <i>Cell, Vol. 129, No. 6. (15 June 2007), pp. 1065-1079.</i><br /><br />Addgene: The karyotypic chaos exhibited by human epithelial cancers complicates efforts to identify mutations critical for malignant transformation. Here we integrate complementary genomic approaches to identify human oncogenes. We show that activation of the ERK and phosphatidylinositol 3-kinase (PI3K) signaling pathways cooperate to transform human cells. Using a library of activated kinases, we identify several kinases that replace PI3K signaling and render cells tumorigenic. Whole genome structural analyses reveal that one of these kinases, IKBKE (IKKepsilon), is amplified and overexpressed in breast cancer cell lines and patient-derived tumors. Suppression of IKKepsilon expression in breast cancer cell lines that harbor IKBKE amplifications induces cell death. IKKepsilon activates the nuclear factor-kappaB (NF-kappaB) pathway in both cell lines and breast cancers. These observations suggest a mechanism for NF-kappaB activation in breast cancer, implicate the NF-kappaB pathway as a downstream mediator of PI3K, and provide a framework for integrated genomic approaches in oncogene discovery. Integrative genomic approaches identify IKBKE as a breast cancer oncogene. JS Boehm JJ Zhao J Yao SY Kim R Firestein IF Dunn SK Sjostrom LA Garraway S Weremowicz AL Richardson H Greulich CJ Stewart LA Mulvey RR Shen L Ambrogio T Hirozane-Kishikawa DE Hill M Vidal M Meyerson JK Grenier G Hinkle DE Root TM Roberts ES Lander K Polyak WC Hahn doi:10.1016/j.cell.2007.03.052 Cell, Vol. 129, No. 6. (15 June 2007), pp. 1065-1079. 2007-09-03T15:15:38-00:00 2007 Cell 0092-8674 129 6 1065 1079 plasmid http://www.citeulike.org/user/jyuh/article/572437 <i>Cell, Vol. 124, No. 6. (24 March 2006), pp. 1283-1298.</i><br /><br />To enable arrayed or pooled loss-of-function screens in a wide range of mammalian cell types, including primary and nondividing cells, we are developing lentiviral short hairpin RNA (shRNA) libraries targeting the human and murine genomes. The libraries currently contain 104,000 vectors, targeting each of 22,000 human and mouse genes with multiple sequence-verified constructs. To test the utility of the library for arrayed screens, we developed a screen based on high-content imaging to identify genes required for mitotic progression in human cancer cells and applied it to an arrayed set of 5,000 unique shRNA-expressing lentiviruses that target 1,028 human genes. The screen identified several known and approximately 100 candidate regulators of mitotic progression and proliferation; the availability of multiple shRNAs targeting the same gene facilitated functional validation of putative hits. This work provides a widely applicable resource for loss-of-function screens, as well as a roadmap for its application to biological discovery. A Lentiviral RNAi Library for Human and Mouse Genes Applied to an Arrayed Viral High-Content Screen. J Moffat DA Grueneberg X Yang SY Kim AM Kloepfer G Hinkle B Piqani TM Eisenhaure B Luo JK Grenier AE Carpenter SY Foo SA Stewart BR Stockwell N Hacohen WC Hahn ES Lander DM Sabatini DE Root doi:10.1016/j.cell.2006.01.040 Cell, Vol. 124, No. 6. (24 March 2006), pp. 1283-1298. 2006-04-01T17:49:23-00:00 2006 Cell 0092-8674 124 6 1283 1298 no-tag