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

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

Research output: Contribution to journal › Article › peer-review

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.

Original language	English (US)
Pages (from-to)	170-174
Number of pages	5
Journal	Molecules and Cells
Volume	23
Issue number	2
State	Published - Apr 30 2007
Externally published	Yes

Keywords

Apoptosis
Automated fluorescence microscope
Hypothetical genes
Subcellular imaging

ASJC Scopus subject areas

Molecular Biology
Cell Biology

Cite this

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title = "Identification of novel regulators of apoptosis using a high-throughput cell-based screen",

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",

author = "Park, {Kyung Mi} and Eunju Kang and Jeon, {Yeo Jin} and Nayoung Kim and Kim, {Nam Soon} and Yoo, {Hyang Sook} and Yeom, {Young Il} and Kim, {Soo Jung}",

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T1 - Identification of novel regulators of apoptosis using a high-throughput cell-based screen

AU - Park, Kyung Mi

AU - Kang, Eunju

AU - Jeon, Yeo Jin

AU - Kim, Nayoung

AU - Kim, Nam Soon

AU - Yoo, Hyang Sook

AU - Yeom, Young Il

AU - Kim, Soo Jung

PY - 2007/4/30

Y1 - 2007/4/30

N2 - 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.

AB - 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.

KW - Apoptosis

KW - Automated fluorescence microscope

KW - Hypothetical genes

KW - Subcellular imaging

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Identification of novel regulators of apoptosis using a high-throughput cell-based screen

Abstract

Keywords

ASJC Scopus subject areas

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