Abstract
Genetically relevant mouse models need to recapitulate the hallmarks of human disease by permitting spatiotemporal gene targeting. This is especially important for replicating the biology of complex diseases like cancer, where genetic events occur in a sporadic fashion within developed somatic tissues. Though a number of renal tubule targeting mouse lines have been developed their utility for the study of renal disease is limited by lack of inducibility and specificity. In this study we describe the generation and characterisation of two novel mouse lines directing CreERT2 expression to renal tubular epithelia. The Pax8-CreERT2 transgenic line uses the mouse Pax8 promoter to direct expression of CreERT2 to all renal tubular compartments (proximal and distal tubules as well as collecting ducts) whilst the Slc22a6-CreERT2 knock-in line utilises the endogenous mouse Slc22a6 locus to specifically target the epithelium of proximal renal tubules. Both lines show high organ and tissue specificity with no extrarenal activity detected. To establish the utility of these lines for the study of renal cancer biology, Pax8-CreERT2 and Slc22a6-CreERT2 mice were crossed to conditional Vhl knockout mice to induce long-term renal tubule specific Vhl deletion. These models exhibited renal specific activation of the hypoxia inducible factor pathway (a VHL target). Our results establish Pax8-CreERT2 and Slc22a6-CreERT2 mice as valuable tools for the investigation and modelling of complex renal biology and disease.
Highlights
Engineered mouse models (GEMM) have been instrumental in understanding the basic principles of renal biology and disease
A Pax8-rtTA transgenic mouse model generated from this locus directs high levels of expression of the reverse tetracyclinedependent transactivator to all proximal, distal and collecting tubules with extrarenal activity occurring only in a minority of hepatocytes [7]
Following pronuclear injection of the 8.4-kb linearized Pax8-CreERT2 construct into C57BL/6 fertilised embryos, we identified five transgenic founders among 17 offspring mice
Summary
Engineered mouse models (GEMM) have been instrumental in understanding the basic principles of renal biology and disease. Though a number of renal tubule targeting mouse lines have been developed, their utility in the study of renal cancer biology is limited by lack of inducibility (Kspcadherin Cre [1], homeobox B7 Cre [2], Aquaporin 2 Cre [3], γ-glutamyl transpeptidase Cre [4], phosphoenolpyruvate carboxykinase Cre [5] and Six homeobox 2 Cre [6]) and/or specificity (phosphoenolpyruvate carboxykinase Cre [5], and Paired box 8 rtTA [7]). Conditional gene expression systems provide an excellent way of controlling gene expression and circumventing any developmental defect as well as early lethality associated with conventional gene targeting These models rely on the use of site-specific recombinases to control the spatiotemporal mutation of the genome. Administration of tamoxifen forces the dissociation of the ERT2-bound heat shock protein 90 (HSP90), allowing CreERT2 to translocate into the nucleus and induce recombination
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