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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
Nov 30, 2023 Vol.46 No.11, pp. 655~725
COVER PICTURE
Kim et al. (pp. 710-724) demonstrated that a pathogen-derived Ralstonia pseudosolanacearum type III effector RipL delays flowering time and enhances susceptibility to bacterial infection in Arabidopsis thaliana. Shown is the RipL-expressing Arabidopsis plant, which displays general dampening of the transcriptional program during pathogen infection, grown in long-day conditions.

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