Strict control of transgene expression in a mouse model for sensitive biological applications based on RMCE compatible ES cells.
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
AbstractRecombinant mouse strains that harbor tightly controlled transgene expression proved to be indispensible tools to elucidate gene function. Different strategies have been employed to achieve controlled induction of the transgene. However, many models are accompanied by a considerable level of basal expression in the non-induced state. Thereby, applications that request tight control of transgene expression, such as the expression of toxic genes and the investigation of immune response to neo antigens are excluded. We developed a new Cre/loxP-based strategy to achieve strict control of transgene expression. This strategy was combined with RMCE (recombinase mediated cassette exchange) that facilitates the targeting of genes into a tagged site in ES cells. The tightness of regulation was confirmed using luciferase as a reporter. The transgene was induced upon breeding these mice to effector animals harboring either the ubiquitous (ROSA26) or liver-specific (Albumin) expression of CreER(T2), and subsequent feeding with Tamoxifen. Making use of RMCE, luciferase was replaced by Ovalbumin antigen. Mice generated from these ES cells were mated with mice expressing liver-specific CreER(T2). The transgenic mice were examined for the establishment of an immune response. They were fully competent to establish an immune response upon hepatocyte specific OVA antigen expression as indicated by a massive liver damage upon Tamoxifen treatment and did not show OVA tolerance. Together, this proves that this strategy supports strict control of transgenes that is even compatible with highly sensitive biological readouts.
CitationStrict control of transgene expression in a mouse model for sensitive biological applications based on RMCE compatible ES cells. 2011, 39 (1):e1 Nucleic Acids Res.
AffiliationHelmholtz Centre for Infection Research, D-38124 Braunschweig, Germany.
JournalNucleic acids research
The following license files are associated with this item:
- A modified RMCE-compatible Rosa26 locus for the expression of transgenes from exogenous promoters.
- Authors: Tchorz JS, Suply T, Ksiazek I, Giachino C, Cloëtta D, Danzer CP, Doll T, Isken A, Lemaistre M, Taylor V, Bettler B, Kinzel B, Mueller M
- Issue date: 2012
- Generation of a mouse line harboring a Bi-transgene expressing luciferase and tamoxifen-activatable creER(T2) recombinase in cartilage.
- Authors: Lo Cascio L, Liu K, Nakamura H, Chu G, Lim NH, Chanalaris A, Saklatvala J, Nagase H, Bou-Gharios G
- Issue date: 2014 Feb
- Ligand-dependent genetic recombination in fibroblasts : a potentially powerful technique for investigating gene function in fibrosis.
- Authors: Zheng B, Zhang Z, Black CM, de Crombrugghe B, Denton CP
- Issue date: 2002 May
- Reproducible doxycycline-inducible transgene expression at specific loci generated by Cre-recombinase mediated cassette exchange.
- Authors: Wong ET, Kolman JL, Li YC, Mesner LD, Hillen W, Berens C, Wahl GM
- Issue date: 2005 Oct 4
- Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells.
- Authors: Nyabi O, Naessens M, Haigh K, Gembarska A, Goossens S, Maetens M, De Clercq S, Drogat B, Haenebalcke L, Bartunkova S, De Vos I, De Craene B, Karimi M, Berx G, Nagy A, Hilson P, Marine JC, Haigh JJ
- Issue date: 2009 Apr