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First inducible transgene expression in porcine large animal models
Author(s) -
Klymiuk Nikolai,
Böcker Wolfgang,
Schönitzer Veronika,
Bahr Andrea,
Radic Tamara,
Fröhlich Thomas,
Wünsch Annegret,
Keßler Barbara,
Kurome Mayuko,
Schilling Eleonore,
Herbach Nadja,
Wanke Rüdiger,
Nagashima Hiroshi,
Mutschler Wolf,
Arnold Georg J.,
Schwinzer Reinhard,
Schieker Matthias,
Wolf Eckhard
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.11-185041
Subject(s) - transgene , transactivation , biology , microbiology and biotechnology , enhancer , gene , expression vector , subcloning , gene expression , activator (genetics) , plasmid , recombinant dna , genetics
The purpose of this study was to establish inducible transgene expression in pigs, a model organism with great promise for experimental physiology and translational medicine, using the binary tet‐on system. This expression system is activated by doxycycline (dox) via the tet‐controlled transactivator (TA). Binding of TA to the transactivator response element (TRE) results in transcription of downstream genes. First, we cloned transgenic founder pigs expressing TA under the control of the CMV enhancer/chicken β‐actin promoter (CAG). Then, cells from CAG‐TA transgenic founders were nucleofected with TRE‐controlled expression vectors for either porcine cytotoxic T‐lymphocyte associated antigen 4‐Fc domain of immunoglobulin G1 (CTLA‐4Ig) or soluble receptor activator of NF‐κB ligand (RANKL), and double‐transgenic offspring were cloned. Dox administration resulted in a dose‐dependent increase in expression of CTLA‐4Ig or RANKL, in nucleofected cells and in transgenic pigs, while in the absence of dox, the levels of both proteins were below the detection limit. Inducible transgene expression was reproduced in double‐transgenic offspring generated by cloning or breeding. Our strategy revealed the first two examples of inducible transgene expression in pigs. The CAG‐TA transgenic pigs generated in this study constitute an interesting basis for future pig models with inducible transgene expression.—Klymiuk, N., Böcker, W., Schönitzer, V., Bahr, A., Radic, T., Fröhlich, T., Wünsch, A., Keßler, B., Kurome, M., Schilling, E., Herbach, N., Wanke, R., Nagashima, H., Mutschler, W., Arnold, G. J., Schwinzer, R., Schieker, M., Wolf, E. First inducible transgene expression in porcine large animal models. FASEB J. 26, 1086‐1099 (2012). www.fasebj.org