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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

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

Article

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

Mol. Cells
May 31, 2023 Vol.46 No.5, pp. 259~328
COVER PICTURE
The alpha-helices in the lamin filaments are depicted as coils, with different subdomains distinguished by various colors. Coil 1a is represented by magenta, coil 1b by yellow, L2 by green, coil 2a by white, coil 2b by brown, stutter by cyan, coil 2c by dark blue, and the lamin Ig-like domain by grey. In the background, cells are displayed, with the cytosol depicted in green and the nucleus in blue (Ahn et al., pp. 309-318).

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