Strict control of transgene expression in a mouse model for sensitive biological applications based on RMCE compatible ES cells.

2.50
HDL Handle:
http://hdl.handle.net/10033/121272
Title:
Strict control of transgene expression in a mouse model for sensitive biological applications based on RMCE compatible ES cells.
Authors:
Sandhu, U; Cebula, M; Behme, S; Riemer, P; Wodarczyk, C; Metzger, D; Reimann, J; Schirmbeck, R; Hauser, H; Wirth, D
Abstract:
Recombinant 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.
Affiliation:
Helmholtz Centre for Infection Research, D-38124 Braunschweig, Germany.
Citation:
Strict 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.
Journal:
Nucleic acids research
Issue Date:
1-Jan-2011
URI:
http://hdl.handle.net/10033/121272
DOI:
10.1093/nar/gkq868
PubMed ID:
20935052
Type:
Article
Language:
en
ISSN:
1362-4962
Appears in Collections:
Publications of AG Modellsysteme für Infektion und Immunität (MSYS)

Full metadata record

DC FieldValueLanguage
dc.contributor.authorSandhu, Uen
dc.contributor.authorCebula, Men
dc.contributor.authorBehme, Sen
dc.contributor.authorRiemer, Pen
dc.contributor.authorWodarczyk, Cen
dc.contributor.authorMetzger, Den
dc.contributor.authorReimann, Jen
dc.contributor.authorSchirmbeck, Ren
dc.contributor.authorHauser, Hen
dc.contributor.authorWirth, Den
dc.date.accessioned2011-02-04T15:20:33Z-
dc.date.available2011-02-04T15:20:33Z-
dc.date.issued2011-01-01-
dc.identifier.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.en
dc.identifier.issn1362-4962-
dc.identifier.pmid20935052-
dc.identifier.doi10.1093/nar/gkq868-
dc.identifier.urihttp://hdl.handle.net/10033/121272-
dc.description.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.en
dc.language.isoenen
dc.titleStrict control of transgene expression in a mouse model for sensitive biological applications based on RMCE compatible ES cells.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for Infection Research, D-38124 Braunschweig, Germany.en
dc.identifier.journalNucleic acids researchen

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