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Nucleic Acids Research, 2001, Vol. 29, No. 7 e39
© 2001 Oxford University Press

A GFP-equipped bidirectional expression module well suited for monitoring tetracycline-regulated gene expression in mouse

H. E. Krestel, M. Mayford1, P. H. Seeburg and R. Sprengel*

Department of Molecular Neurobiology, Max-Planck-Institute for Medical Research, Jahnstrasse. 29, D-69120 Heidelberg, Germany and 1Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037, USA

Doxycycline (Dox)-sensitive co-regulation of two transcriptionally coupled transgenes was investigated in the mouse. For this, we generated four independent mouse lines carrying coding regions for green fluorescent protein (GFP) and ß-galactosidase in a bicistronic, bidirectional module. In all four lines the expression module was silent but was activated when transcription factor tTA was provided by the {alpha}-CaMKII-tTA transgene. In vivo analysis of GFP fluorescence, ß-galactosidase and immunochemical stainings revealed differences in GFP and ß-galactosidase levels between the lines, but comparable patterns of expression. Strong signals were found in neurons of the olfactory system, neocortical, limbic lobe and basal ganglia structures. Weaker expression was limited to thalamic, pontine and medullary structures, the spinal cord, the eye and to some Purkinje cells in the cerebellum. Strong GFP signals were always accompanied by intense ß-galactosidase activity, both of which could be co-regulated by Dox. We conclude that the tTA-sensitive bidirectional expression module is well suited to express genes of interest in a regulated manner and that GFP can be used to track transcriptional activity of the module in the living mouse.

* To whom correspondence should be addressed. Tel: +49 6221 486101; Fax: +49 6221 486110; Email: sprengel{at}mpimf-heidelberg.mpg.de


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