The overall goal of Core B is to support this POI in all aspects involving RNAi-based knock-down technology. The participating Projects rely to a great extent on cellular systems as platforms for discovery, as well as on our ability to assess gene-function in them. In the absence of genetic approaches for gene ablation in somatic cells, RNAi is the ideal tool for such purpose as a rapid, efficient, inexpensive, versatile, and easy to use method for gene knock-down. Although RNAi approaches also present some challenges, the Core will put in place working strategies to overcome them.
Our first aim will be to generate multiple, high-efficiency RNAi-reagents to inhibit the expression of targets that we expect will arise in the course of this PPG as potential regulators of the PTEN-Siah2 pathway. These reagents will be instrumental in evaluating the potential of such regulators as therapeutic targets for the treatment of cancer types derived from PTEN deficiency. To achieve this goal we will systematically generate multiple lentiviral-shRNAs and empirically assess their knock-down potency. The selected reagents will be transduced into the various cell lines and transferred to project teams at various stages of this program. Our second focus will be siRNA screening, as an unbiased tactic to discover new biology surrounding PTEN related tumorogenesis and pinpoint new potential targets. To accomplish these goals, we will provide RNAi libraries, state of the art equipment, expertise and manpower to develop high throughput cell-based assays, execute screens, analyze and confirm results, and produce optimized reagents for follow-up studies. Core B will interact closely with the 3 Projects and Core C, to which it will be providing reagents and services.
|Maruyama, Takeshi; Araki, Toshihiro; Kawarazaki, Yosuke et al. (2014) Roquin-2 promotes ubiquitin-mediated degradation of ASK1 to regulate stress responses. Sci Signal 7:ra8|
|Kim, H; Claps, G; Moller, A et al. (2014) Siah2 regulates tight junction integrity and cell polarity through control of ASPP2 stability. Oncogene 33:2004-10|
|Scortegagna, Marzia; Kim, Hyungsoo; Li, Jian-Liang et al. (2014) Fine tuning of the UPR by the ubiquitin ligases Siah1/2. PLoS Genet 10:e1004348|
|Qi, Jianfei; Kim, Hyungsoo; Scortegagna, Marzia et al. (2013) Regulators and effectors of Siah ubiquitin ligases. Cell Biochem Biophys 67:15-24|
|Feng, Yongmei; Lau, Eric; Scortegagna, Marzia et al. (2013) Inhibition of melanoma development in the Nras((Q61K)) ::Ink4a(-/-) mouse model by the small molecule BI-69A11. Pigment Cell Melanoma Res 26:136-42|
|Barile, Elisa; De, Surya K; Feng, Yongmei et al. (2013) Synthesis and SAR studies of dual AKT/NF-*B inhibitors against melanoma. Chem Biol Drug Des 82:520-33|
|Stebbins, John L; Santelli, Eugenio; Feng, Yongmei et al. (2013) Structure-based design of covalent Siah inhibitors. Chem Biol 20:973-82|
|Filipp, Fabian V; Scott, David A; Ronai, Ze'ev A et al. (2012) Reverse TCA cycle flux through isocitrate dehydrogenases 1 and 2 is required for lipogenesis in hypoxic melanoma cells. Pigment Cell Melanoma Res 25:375-83|
|Feng, Yongmei; Barile, Elisa; De, Surya K et al. (2011) Effective inhibition of melanoma by BI-69A11 is mediated by dual targeting of the AKT and NF-*B pathways. Pigment Cell Melanoma Res 24:703-13|
|Scott, David A; Richardson, Adam D; Filipp, Fabian V et al. (2011) Comparative metabolic flux profiling of melanoma cell lines: beyond the Warburg effect. J Biol Chem 286:42626-34|
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