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1 Department of Medicine, University of Utah, Salt Lake City, UT, USA
2 Departments of Adult and Pediatric, University of Utah, Salt Lake City, UT, USA
3 Department of Medicine, University of Utah, Salt Lake City, UT, USA; Veterans Affairs Medical Center, Salt Lake City, UT, USA
* To whom correspondence should be addressed. E-mail: donald.kohan{at}hsc.utah.edu.
Evaluation of thick ascending limb (TAL) function has been hindered by the limited ability to selectively examine the function of this nephron segment in vivo. To address this, a Cre/loxP strategy was employed whereby the Tamm-Horsfall (THP) promoter was used to drive Cre recombinase expression in transgenic mice. The THP gene was cloned from a mouse genomic library and 3.7 kb of the mouse THP 5'-flanking region containing the 1st non-coding exon of the THP gene was inserted upstream of an epitope-tagged Cre recombinase (THP-CreTag). THP-CreTag transgenic mice were bred with ROSA26-eYFP mice (contain a loxP-flanked "STOP" sequence 5' to enhanced yellow fluorescent protein (eYFP)) and doubly heterozygous offspring analyzed. THP and eYFP were expressed in an identical pattern with predominant localization to the renal outer medulla without expression in non-renal tissues. eYFP did not co-localize with thiazide-sensitive cotransporter (distal tubule) or neuronal nitric oxide synthase (macula densa) expression. THP mRNA expression was detected only in kidney, while CreTag mRNA was also present in testes. These data indicate that THP-CreTag transgenic mice can be used to TAL-specific gene recombination in the kidney.
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