Please use this identifier to cite or link to this item: https://doi.org/10.1002/bit.22152
Title: Mutated polyadenylation signals for controlling expression levels of multiple genes in mammalian cells
Authors: Yang, Y.
Mariati
Ho, S.C.L.
Yap, M.G.S. 
Keywords: Control of gene expression
Mammalian cells
Monoclonal antibody
RNA processing
SV40 early polyadenylation signal
Issue Date: 1-Mar-2009
Citation: Yang, Y., Mariati, Ho, S.C.L., Yap, M.G.S. (2009-03-01). Mutated polyadenylation signals for controlling expression levels of multiple genes in mammalian cells. Biotechnology and Bioengineering 102 (4) : 1152-1160. ScholarBank@NUS Repository. https://doi.org/10.1002/bit.22152
Abstract: A set of mutated SV40 early polyadenylation signals (SV40pA) with varying strengths is generated by mutating the AATAAA sequence in the wild-type SV40pA. They are shown to control the expression level ofa gene over a 10-fold range using luciferase reporter genes in transient transfection assays. The relative strength of these SV40pA variants remains similar under three commonly used mammalian promoters and in five mammalian cell lines. Application of SV40pA variants for controlling expression level of multiple genes is demonstrated in a study of monoclonal antibody (mAb) synthesis in mammalian cells. By using SV40pA variants of different strengths, the expression of light chain (LC) and heavy chain (HC) genes encoded in a single vector is independently altered which results in different ratios of LC to HC expression spanning a range from 0.24 to 16.42. The changes in gene expression are determined by measuring mRNA levels and intracellular LC and HC polypeptides. It is found that a substantial decrease of HC expression, which increases the LC/HC mRNA ratio, only slightly reduces mAb production. However, reducing the LC expression by a similar magnitude, which decreases the LC/HC mRNA ratio results in a sharp decline of mAb production to trace amounts. This set of SV40pA variants offers a new tool for accurate control of the relative expression levels of multiple genes. It will have wide-ranging applications in fields related to the study of biosynthesis of multi-subunit proteins, proteomic research on protein interactions, and multi-gene metabolic engineering. © 2008 Wiley Periodicals, Inc.
Source Title: Biotechnology and Bioengineering
URI: http://scholarbank.nus.edu.sg/handle/10635/89542
ISSN: 00063592
DOI: 10.1002/bit.22152
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

24
checked on Jun 14, 2018

WEB OF SCIENCETM
Citations

20
checked on May 9, 2018

Page view(s)

25
checked on May 11, 2018

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.