Large-scale loss of function RNAi screening has the greatest potential for the discovery of novel gene function and the identification of new protein targets with clinical relevance. Building upon our experience using synthetic siRNAs to down-regulate individual genes (ZO1 BC 010613) we have established robust RNAi screening formats using synthetic siRNAs. Developing an RNAi screening capability has required extensive optimization of the induction of RNAi by synthetic siRNAs in a manner compatible with a larger scale workflow that generates robust and reproducible data. Critical to establishing effective RNAi screening has been the development of optimized protocols, statistical analysis, and down-stream validation procedures and this enables us to screen several thousand genes simultaneously. Our independent and collaborative studies have established conditions for synthetic siRNA-mediated RNAi screening in several cancer cell lines, including those used for the study of breast, ovarian, colorectal cancer, multiple myeloma and Ewing's Sarcoma. Independent and collaborative RNAi screens to identify novel cancer-associated genes, including genes that can be exploited directly as anti-cancer molecular targets have been initiated. This hypothesis-generating approach has enabled us to identify a number of proteins that influence the growth of cancer cell lines and follow-up analysis of these specific proteins is on-going.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010615-09
Application #
8552749
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
9
Fiscal Year
2012
Total Cost
$337,555
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Garimella, Sireesha V; Gehlhaus, Kristie; Dine, Jennifer L et al. (2014) Identification of novel molecular regulators of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in breast cancer cells by RNAi screening. Breast Cancer Res 16:R41
Camps, Jordi; Pitt, Jason J; Emons, Georg et al. (2013) Genetic amplification of the NOTCH modulator LNX2 upregulates the WNT/β-catenin pathway in colorectal cancer. Cancer Res 73:2003-13
Tandle, Anita T; Kramp, Tamalee; Kil, Whoon J et al. (2013) Inhibition of polo-like kinase 1 in glioblastoma multiforme induces mitotic catastrophe and enhances radiosensitisation. Eur J Cancer 49:3020-8
Hummon, Amanda B; Pitt, Jason J; Camps, Jordi et al. (2012) Systems-wide RNAi analysis of CASP8AP2/FLASH shows transcriptional deregulation of the replication-dependent histone genes and extensive effects on the transcriptome of colorectal cancer cells. Mol Cancer 11:1
Bennett, Christina N; Tomlinson, Christine C; Michalowski, Aleksandra M et al. (2012) Cross-species genomic and functional analyses identify a combination therapy using a CHK1 inhibitor and a ribonucleotide reductase inhibitor to treat triple-negative breast cancer. Breast Cancer Res 14:R109
Martin, S E; Wu, Z-H; Gehlhaus, K et al. (2011) RNAi screening identifies TAK1 as a potential target for the enhanced efficacy of topoisomerase inhibitors. Curr Cancer Drug Targets 11:976-86
Murrow, Lyndsay M; Garimella, Sireesha V; Jones, Tamara L et al. (2010) Identification of WEE1 as a potential molecular target in cancer cells by RNAi screening of the human tyrosine kinome. Breast Cancer Res Treat 122:347-57