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Mol. Cells 2007; 23(2): 170-174
Published online January 1, 1970
© The Korean Society for Molecular and Cellular Biology
Identification of Novel Regulators of Apoptosis Using a High-Throughput Cell-based Screen
High-throughput subcellular imaging is a powerful tool for investigating the function of genes. In order to identify novel regulators of apoptosis we transiently transfected HeLa cells with 938 hypothetical genes of unknown function, and captured their nuclear images with an automated fluorescence microscope. We selected genes that induced greater than 3-fold increase in the percentage of apoptotic nuclei compared with vector-transfected cells. The full-length genes C10orf61, MGC 26717, and FLJ13855 were identified as candidate proapoptotic genes, and their apoptotic effects were confirmed by DNA fragmentation ELISAs and Western blotting for caspase-7 and PARP. We conclude that a subcellular image-based apoptotic screen is useful for identifying genes with proapoptotic activity.
Keywords Apoptosis; Automated Fluorescence Microscope;, Hypothetical Genes; Subcellular Imaging
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Research Article
Mol. Cells 2007; 23(2): 170-174
Published online April 30, 2007
Copyright © The Korean Society for Molecular and Cellular Biology.
Identification of Novel Regulators of Apoptosis Using a High-Throughput Cell-based Screen
Kyung Mi Park, Eunju Kang, Yeo-Jin Jeon, Nayoung Kim, Nam-Soon Kim, Hyang-Sook Yoo, Young Il Yeom, Soo Jung Kim
Abstract
High-throughput subcellular imaging is a powerful tool for investigating the function of genes. In order to identify novel regulators of apoptosis we transiently transfected HeLa cells with 938 hypothetical genes of unknown function, and captured their nuclear images with an automated fluorescence microscope. We selected genes that induced greater than 3-fold increase in the percentage of apoptotic nuclei compared with vector-transfected cells. The full-length genes C10orf61, MGC 26717, and FLJ13855 were identified as candidate proapoptotic genes, and their apoptotic effects were confirmed by DNA fragmentation ELISAs and Western blotting for caspase-7 and PARP. We conclude that a subcellular image-based apoptotic screen is useful for identifying genes with proapoptotic activity.
Keywords: Apoptosis, Automated Fluorescence Microscope,, Hypothetical Genes, Subcellular Imaging
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