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.
Park
Kyung Mi
KM
Functional Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Korea.
Kang
Eunju
E
Jeon
Yeo-Jin
YJ
Kim
Nayoung
N
Kim
Nam-Soon
NS
Yoo
Hyang-Sook
HS
Yeom
Young Il
YI
Kim
Soo Jung
SJ
D017209:Apoptosis / Q000502:physiology*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D053179:Caspase 7 / Q000378:metabolism; D024161:Collagen Type XI / Q000378:metabolism; D053938:DNA Fragmentation; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism
D051017:Apoptosis Regulatory Proteins; C499319:COL11A2 protein, human; D024161:Collagen Type XI; D053179:Caspase 7