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Acta Physiologica Congress

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Acta Physiologica 2011; Volume 203, Supplement 686
Joint Congress of FEPS and Turkish Society of Physiological Sciences
9/3/2011-9/7/2011
Istanbul, Turkey


HIGH THROUGHPUT EXPERIMENTATION FOR HYPOTHESIS GENERATION AND TESTING
Abstract number: W23

Chan1 Ting, Ming1 Jie, Chao1 Pang, Demiralp2 Duygu Ozel, Aktas1 Bertal Huseyin

1Harvard Medical School, Boston, MA, USA
2Ankara University, Biotechnology Institute, Proteomic Unit of the Central Laboratory Ankara, Turkey

Classical cell/molecular biology approach of studying one or a few molecules at a time for their role in normal/patho-biology limits the speed of scientific discovery and often lead to conclusions with narrow applicability. The alternative of studying all genes/proteins for their role in normal- and patho-biology circumvents these limitations and allows for identification of dominant and secondary functional genetic networks and generation of novel hypothesis that may be tested using similar system-wide approaches. The experimental system should have high throughput, be highly accurate, reproducible, and cost effective. An example of high-throughput hypothesis generation is our studies of the molecular and chemical genetics of the integrated endoplasmic reticulum (ER) stress response (IERSR). The ER is the site of protein folding and processing in the cells. Our knowledge of how ER homeostasis is maintained, and if/how disturbances in ER homeostasis contribute to patho-biology of human disorders is poorly understood. To ameliorate this, we developed a high throughput experimental system capable of high-throughput genetic and chemical interrogation of ER-physiology. We generated two dual-luciferase assay systems to interrogate two different arms of IERSR, the PERK/eIF2a arm that reduced protein synthesis, and Ire1/Xbp-1 arm that increases size and folding capacity of the ER to resolve the ER-stress. We will show how we developed these assay systems, how we screen siRNA, micro-RNA, morpholino and small chemical libraries, how we integrate data from two reporters in two different assay systems and genetic and chemical screens to generate and test new hypotheses. Finally we will demonstrate how this new approach is changing the face of the scientific inquiry.

To cite this abstract, please use the following information:
Acta Physiologica 2011; Volume 203, Supplement 686 :W23

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