Abstract
The Cre/loxP recombination system has revolutionized the ability to genetically manipulate animal genomes in order to conditionally control gene expression. With recent advances in genome editing, barriers to manipulating the rat genome have been overcome and it is now possible to generate new rat strains (Cre drivers) in which Cre recombinase expression is carefully controlled temporally and/or spatially. However, the ability to evaluate and characterize these Cre driver strains is limited by the availability of reliable reporter rat strains. Here, we describe the generation and characterization of a new transgenic rat strain in which conditional expression of the ZsGreen fluorescent protein gene requires the presence of exogenous Cre recombinase. Breeding Cre-expressing rat strains to this stable ZsGreen reporter strain provides an ideal method for validating new rat Cre driver lines and will greatly accelerate the characterization pipeline.
Highlights
The Cre/loxP recombination system has revolutionized the ability to genetically manipulate animal genomes in order to spatially and temporally control gene expression[1]
We describe the generation and characterization of a new transgenic rat strain in which conditional expression of the ZsGreen fluorescent protein gene requires the presence of exogenous Cre recombinase
Breeding Cre-expressing rat strains to this stable ZsGreen reporter strain provides an ideal method for validating new rat Cre driver lines
Summary
The Cre/loxP recombination system has revolutionized the ability to genetically manipulate animal genomes in order to spatially and temporally control gene expression[1] This has resulted in the generation of a large number of mouse strains, often called “Cre drivers”, in which Cre recombinase expression is carefully controlled. The strains would need to be cryorecovered in order for an investigator to use them, resulting in a delay in the availability of live animals Cre-mediated recombination results in deletion of the EGFP gene with subsequent expression of mCherry instead[2] This strain is available currently as live animals at the Rat Resource in China. We show that the ZsGreen transgene continues to exhibit stable expression in the presence of Cre recombinase even in reporter animals multiple generations removed from the original founder animal, demonstrating that transgene silencing has not occurred over time as has been seen in other transgenic reporter strains
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