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Proc Natl Acad Sci U S A. 2013 Mar 26;110(13):5004-9. doi: 10.1073/pnas.1218620110. Epub 2013 Mar 8.

Simple piggyBac transposon-based mammalian cell expression system for inducible protein production.

Author information

1
Department of Biochemistry, University of Toronto, Toronto, ON, Canada M5S 1A8.

Abstract

Reported here is a piggyBac transposon-based expression system for the generation of doxycycline-inducible, stably transfected mammalian cell cultures for large-scale protein production. The system works with commonly used adherent and suspension-adapted mammalian cell lines and requires only a single transfection step. Moreover, the high uniform expression levels observed among clones allow for the use of stable bulk cell cultures, thereby eliminating time-consuming cloning steps. Under continuous doxycycline induction, protein expression levels have been shown to be stable for at least 2 mo in the absence of drug selection. The high efficiency of the system also allows for the generation of stable bulk cell cultures in 96-well format, a capability leading to the possibility of generating stable cell cultures for entire families of membrane or secreted proteins. Finally, we demonstrate the utility of the system through the large-scale production (140-750 mg scale) of an endoplasmic reticulum-resident fucosyltransferase and two potential anticancer protein therapeutic agents.

PMID:
23476064
PMCID:
PMC3612669
DOI:
10.1073/pnas.1218620110
[Indexed for MEDLINE]
Free PMC Article

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