Abstract
BackgroundTurning gene expression on and off at will is one of the most powerful tools for the study of gene function in vivo. While several conditional systems were successful in invertebrates, in mice the Cre/loxP recombination system and the tet-controlled transcription activation system are predominant. Both expression systems allow for spatial and temporal control of gene activities, and, in the case of tet regulation, even for the reversible activation/inactivation of gene expression. Although the rat is the principal experimental model in biomedical research, in particular in studies of neuroscience, conditional rat transgenic systems are exceptionally rare in this species.ResultsWe addressed this lack of technology, and established and thoroughly characterized CreERT2 and tTA transgenic rats with forebrain-specific transgene expression, controlled by the CaMKII alpha promoter. In addition, we developed new universal rat reporter lines for both transcription control systems and established inducible and efficient reporter gene expression in forebrain neurons.ConclusionsWe demonstrate that conditional genetic manipulations in the rat brain are both feasible and practicable and outline advantages and limitations of the Tet and Cre/loxP system in the rat brain.
Highlights
Turning gene expression on and off at will is one of the most powerful tools for the study of gene function in vivo
We describe the generation of transgenic rat lines expressing tTA and the tamoxifen-inducible Cre recombinase CreERT2 under the control of the forebrainspecific Ca2+/calmodulin-dependent protein kinase IIa (CaMKIIa) promoter
Such animals will express the transgene depending on administration of the tet derivative doxycycline hydrochloride (Dox), which can be supplied in the drinking water
Summary
Turning gene expression on and off at will is one of the most powerful tools for the study of gene function in vivo. The most popular approaches for the generation of genetically modified mice are the targeted engineering of genomic DNA in embryonic stem cells to introduce gene knockouts and the DNA microinjection into the pronuclei of fertilized eggs, which results in random integration of the transgene into the genome for the overexpression of gene products. These valuable techniques have inherent limitations related to the irreversibility and ubiquity of the germ line genetic modifications. Once the ligand tamoxifen is applied, CreERT2 translocates into the nucleus, where recombination takes place
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