Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
AbstractLong-term, recombinant gene expression in mammalian cells depends on the nature of the transgene integration site and its inherent properties to modulate transcription (epigenetic effects). Here we describe a method by which high transgene expression is achieved and stabilized in extensively proliferating cultures. The method is based on strict co-expression of the transgene with an antitoxin in cells that express the respective toxin. Since the strength of antitoxin expression correlates with an advantage for cell growth, the cells with strong antitoxin expression are enriched over time in cultures of heterogeneous cells. This principle was applied to CHO cell lines that conditionally express the toxin kid and that are transduced to co-express the antitoxin kis together with different transgenes of interest. Cultivation of pools of transfectants that express the toxin steadily increase their transgene expression within several weeks to reach a maximum that is up to 120-fold over the initial status. In contrast, average transgene expression drops in the absence of toxin expression. Together, we show that cells conditionally expressing kid can be employed to create overexpressing cells by a simple coupling of kis to the transgene of interest, without further manipulation and in absence of selectable drugs.
CitationToxin-antitoxin based transgene expression in mammalian cells. 2010, 38 (5):e32 Nucleic Acids Res.
AffiliationHelmholtz Centre for Infection Research, Braunschweig, Germany.
JournalNucleic acids research
The following license files are associated with this item:
- Interactions of Kid-Kis toxin-antitoxin complexes with the parD operator-promoter region of plasmid R1 are piloted by the Kis antitoxin and tuned by the stoichiometry of Kid-Kis oligomers.
- Authors: Monti MC, Hernández-Arriaga AM, Kamphuis MB, López-Villarejo J, Heck AJ, Boelens R, Díaz-Orejas R, van den Heuvel RH
- Issue date: 2007
- Cleavage of the antitoxin of the parD toxin-antitoxin system is determined by the ClpAP protease and is modulated by the relative ratio of the toxin and the antitoxin.
- Authors: Diago-Navarro E, Hernández-Arriaga AM, Kubik S, Konieczny I, Díaz-Orejas R
- Issue date: 2013 Jul
- Enhanced transgene expression using cis-acting elements combined with the EF1 promoter in a mammalian expression system.
- Authors: Wang W, Guo X, Li YM, Wang XY, Yang XJ, Wang YF, Wang TY
- Issue date: 2018 Oct 15
- RNase/anti-RNase activities of the bacterial parD toxin-antitoxin system.
- Authors: Muñoz-Gómez AJ, Lemonnier M, Santos-Sierra S, Berzal-Herranz A, Díaz-Orejas R
- Issue date: 2005 May
- Interactions between the toxin Kid of the bacterial parD system and the antitoxins Kis and MazE.
- Authors: Kamphuis MB, Monti MC, van den Heuvel RH, Santos-Sierra S, Folkers GE, Lemonnier M, Díaz-Orejas R, Heck AJ, Boelens R
- Issue date: 2007 Apr 1