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dc.contributor.authorKoh, Esther YCen
dc.contributor.authorYeo, Jessna HMen
dc.contributor.authorHo, Steven CLen
dc.contributor.authorYang, Yuanshengen
dc.identifier.citationMariati, Koh, E. Y., Yeo, J. H., Ho, S. C., & Yang, Y. (2014). Toward stable gene expression in CHO cells. Bioengineered, 5(5), 340-345.en
dc.description.abstractMaintaining 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.en
dc.description.sponsorshipASTAR (Agency for Sci., Tech. and Research, S’pore)en
dc.format.extent6 p.en
dc.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: [].en
dc.subjectcore CpG island elementen
dc.subjectgene silencingen
dc.subjectexpression stabilityen
dc.titleToward stable gene expression in CHO cells Preventing promoter silencing with core CpG island elementsen
dc.typeJournal Articleen
dc.contributor.schoolSchool of Chemical and Biomedical Engineeringen
dc.description.versionAccepted versionen
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