Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/81465
Title: Impact of Using Different Promoters and Matrix Attachment Regions on Recombinant Protein Expression Level and Stability in Stably Transfected CHO Cells
Authors: Ho, Steven C. L.
Mariati
Yeo, Jessna H. M.
Fang, Shiyi Goh
Yang, Yuansheng
Keywords: Matrix attachment region (MAR)
CMV promoter
SV40 promoter
CHO elongation factor 1α promoter
Stable recombinant protein expression
CHO cells
Issue Date: 2014
Source: Ho, S. C. L., Mariati, Yeo, J. H. M., Fang, S. G., & Yang, Y. (2015). Impact of Using Different Promoters and Matrix Attachment Regions on Recombinant Protein Expression Level and Stability in Stably Transfected CHO Cells. Molecular Biotechnology, 57(2), 138-144.
Series/Report no.: Molecular Biotechnology
Abstract: High expression level and long-term expression stability are required for therapeutic protein production in mammalian cells. Three commonly used promoters from the simian virus 40 (SV40), the CHO elongation factor 1α gene (EF1α), and the human cytomegalovirus major immediate early gene (CMV) and two matrix attachment regions from the chicken lysozyme gene (cMAR) and the human interferon β (iMAR) were evaluated for enhancing recombinant gene expression level and stability in stably transfected CHO cells. In the absence of MAR elements, the SV40 promoter gave lower expression level but higher stability than the EF1α promoter and the CMV promoter. The inclusion of MAR elements did not increase the integrated gene copies for all promoters but did enhance expression level for only the SV40 promoter. The enhanced gene expression was due to an increase in mRNA levels. Neither MAR elements enhance gene expression stability during long-term culture. The combinations of SV40 promoter and MAR elements are the best for obtaining both high expression level and stability. The information presented here would be valuable to those developing vectors for generation of CHO cell lines with stable and high productivity.
URI: https://hdl.handle.net/10356/81465
http://hdl.handle.net/10220/40789
ISSN: 1073-6085
DOI: 10.1007/s12033-014-9809-2
Rights: © 2014 Springer Science+Business Media New York. This is the author created version of a work that has been peer reviewed and accepted for publication by Molecular Biotechnology, Springer Science+Business Media New York. 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/10.1007/s12033-014-9809-2].
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SCBE Journal Articles

SCOPUSTM   
Citations 10

46
Updated on Dec 2, 2022

Web of ScienceTM
Citations 10

42
Updated on Dec 4, 2022

Page view(s)

248
Updated on Dec 7, 2022

Download(s) 10

306
Updated on Dec 7, 2022

Google ScholarTM

Check

Altmetric


Plumx

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