Amplified dicistronic expression units mediate apoptosis protection in CHO-DG44 cells adapted for growth in serum-free media: impact on mitochondria copy number
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Date
2005Type
- Conference Paper
ETH Bibliography
yes
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Abstract
We have constructed dicistronic expression vectors for one-step antiapoptosis engineering and product expression. Expression units encoding the soluble intercellular adhesion molecule 1 (sICAM), a potential therapeutic for treatment of the common cold, and bcl-2/bcl-xL were integrated and amplified in Chinese hamster ovary- (CHO) DG44 cells adapted for anchorage-independent growth in serum-free media. sICAM expression was translated in a cap-dependent and survival gene expression in a cap-independent manner based on the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES). Batch cultivations of engineered CHO-DG44 cells containing amplified transgene expression units exhibited improved viability and delayed onset of apoptosis compared to cell lines harboring monocistronic control constructs. bcl-xL-mediated apoptosis protection was significantly higher compared to bcl-2-based survival engineering. High-level expression of bcl-2 and bcl-xL seem to be required to compensate for increased mitochondria numbers found to be associated with production cell lines grown in serum-free medium. Show more
Publication status
publishedExternal links
Book title
Animal Cell Technology Meets GenomicsJournal / series
ESACT ProceedingsVolume
Pages / Article No.
Publisher
SpringerEvent
Organisational unit
03694 - Fussenegger, Martin / Fussenegger, Martin
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Is part of: http://hdl.handle.net/20.500.11850/694418
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