Mol. Cells 2009; 27(5): 577-582
Published online May 15, 2009
https://doi.org/10.1007/s10059-009-0078-z
© The Korean Society for Molecular and Cellular Biology
The development of a high-throughput functional ge-nomic screening provides a novel and expeditious ap-proach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regu-lates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon ?, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.
Keywords HEK293, HUGT1, monoclonal antibody, recombinant protein production, unfolded protein response
Mol. Cells 2009; 27(5): 577-582
Published online May 31, 2009 https://doi.org/10.1007/s10059-009-0078-z
Copyright © The Korean Society for Molecular and Cellular Biology.
Sebastian Chih Yuan Ku, Teng Rhui Lwa, Maybelline Giam, Miranda Gek Sim Yap, and Sheng-Hao Chao
The development of a high-throughput functional ge-nomic screening provides a novel and expeditious ap-proach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regu-lates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon ?, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.
Keywords: HEK293, HUGT1, monoclonal antibody, recombinant protein production, unfolded protein response
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