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research article

Development of stable cell lines for production or regulated expression using matrix attachment regions

Zahn-Zabal, M.
•
Kobr, M.
•
Girod, P. A.
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2001
Journal of biotechnology

One of the major hurdles of isolating stable, inducible or constitutive high-level producer cell lines is the time-consuming selection procedure. Given the variation in the expression levels of the same construct in individual clones, hundreds of clones must be isolated and tested to identify one or more with the desired characteristics. Various boundary elements (BEs), matrix attachment regions, and locus control regions (LCRs) were screened for their ability to augment the expression of heterologous genes in Chinese hamster ovary (CHO) cells. Of the chromatin elements assayed, the chicken lysozyme matrix-attachment region (MAR) was the only element to significantly increase stable reporter expression. We found that the use of the MAR increases the proportion of high-producing clones, thus reducing the number of clones that need to be screened. These benefits are observed both for constructs with MARs flanking the transgene expression cassette, as well as when constructs are co-transfected with the MAR on a separate plasmid. Moreover, the MAR was co-transfected with a multicomponent regulatable beta-galactosidase expression system in C2C12 cells and several clones exhibiting regulated expression were identified. Hence, MARs are useful in the development of stable cell lines for production or regulated expression.

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Type
research article
DOI
10.1016/S0168-1656(00)00423-5
Web of Science ID

WOS:000168113000003

PubMed ID

11267697

Author(s)
Zahn-Zabal, M.
•
Kobr, M.
•
Girod, P. A.
•
Imhof, M.
•
Chatellard, P.
•
de Jesus, M.  
•
Wurm, F.  
•
Mermod, N.
Date Issued

2001

Published in
Journal of biotechnology
Volume

87

Issue

1

Start page

29

End page

42

Subjects

Animals

•

*CHO Cells

•

Cell Line

•

Chickens

•

Chromatin/genetics

•

Cricetinae

•

Extracellular Matrix/*metabolism

•

Gene Expression Regulation

•

Muramidase/*genetics/metabolism

•

Protein Engineering/*methods

•

Transfection

•

Transgenes

Note

Laboratory of Molecular Biotechnology, Center for Biotechnology UNIL-EPFL, University of Lausanne, CBUE, DC-IGC, CH-1015, Lausanne, Switzerland.

Evaluation Studies

Journal Article

Research Support, Non-U.S. Gov't

Netherlands

Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LBTC  
Available on Infoscience
June 5, 2007
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/7623
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