title: Simple piggyBac transposon-based mammalian cell expression system for inducible protein production.
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.
authors.affiliation: Department of Biochemistry, University of Toronto, Toronto, ON, Canada M5S 1A8.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Li
authors.forename: Iacovos P
authors.initials: IP
authors.lastname: Michael
authors.forename: Dongxia
authors.initials: D
authors.lastname: Zhou
authors.forename: Andras
authors.initials: A
authors.lastname: Nagy
authors.forename: James M
authors.initials: JM
authors.lastname: Rini
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004251:DNA Transposable Elements; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis*; D020218:Response Elements; D013752:Tetracycline / Q000494:pharmacology; D014162:Transfection
chemical_list: D000900:Anti-Bacterial Agents; D004251:DNA Transposable Elements; D011994:Recombinant Proteins; D013752:Tetracycline