Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/81451
Title: Toward stable gene expression in CHO cells Preventing promoter silencing with core CpG island elements
Authors: Mariati
Koh, Esther YC
Yeo, Jessna HM
Ho, Steven CL
Yang, Yuansheng
Keywords: CHO
core CpG island element
gene silencing
expression stability
Issue Date: 2014
Source: Mariati, Koh, E. Y., Yeo, J. H., Ho, S. C., & Yang, Y. (2014). Toward stable gene expression in CHO cells. Bioengineered, 5(5), 340-345.
Series/Report no.: Bioengineered
Abstract: Maintaining high gene expression level during long-term culture is critical when producing therapeutic recombinant proteins using mammalian cells. Transcriptional silencing of promoters, most likely due to epigenetic events such as DNA methylation and histone modifications, is one of the major mechanisms causing production instability. Previous studies demonstrated that the core CpG island element (IE) from the hamster adenine phosphoribosyltransferase gene is effective to prevent DNA methylation. We generated one set of modified human cytomegalovirus (hCMV) promoters by insertion of one or two copies of IE in either forward or reverse orientations into different locations of the hCMV promoter. The modified hCMV with one copy of IE inserted between the hCMV enhancer and core promoter in reverse orientation (MR1) was most effective at enhancing expression stability in CHO cells without comprising expression level when compared with the wild type hCMV. We also found that insertion of IE into a chimeric murine CMV (mCMV) enhancer and human elongation factor-1α core (hEF) promoter in reverse orientation did not enhance expression stability, indicating that the effect of IE on expression stability is possibly promoter specific.
URI: https://hdl.handle.net/10356/81451
http://hdl.handle.net/10220/40784
ISSN: 1949-1018
DOI: 10.4161/bioe.32111
Rights: © 2014 Landes Bioscience. This is the author created version of a work that has been peer reviewed and accepted for publication in Bioengineered, published by Taylor & Francis on behalf of Landes Bioscience. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document.  The published version is available at: [http://dx.doi.org/10.4161/bioe.32111].
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SCBE Journal Articles

Files in This Item:
File Description SizeFormat 
Toward stable gene expression in CHO cells.pdf339.07 kBAdobe PDFThumbnail
View/Open

SCOPUSTM   
Citations 20

14
checked on Sep 5, 2020

WEB OF SCIENCETM
Citations 20

14
checked on Oct 24, 2020

Page view(s) 20

123
checked on Oct 25, 2020

Download(s) 20

94
checked on Oct 25, 2020

Google ScholarTM

Check

Altmetric


Plumx

Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.