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Physiol. Genomics 2: 67-75, 2000;
1094-8341/00 $5.00
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Received 3 November 1999; accepted in final form 28 January 2000.
Physiological Genomics 2:67-75 (2000)
1094-8341/00 $5.00 © 2000 American Physiological Society

Targeting the Hprt locus in mice reveals differential regulation of Tie2 gene expression in the endothelium

VALERIE EVANS1, ANTONIS HATZOPOULOS2, WILLIAM C. AIRD3, HELEN B. RAYBURN1, ROBERT D. ROSENBERG1 and JAN ALBERT KUIVENHOVEN4

1 Massachusetts Institute of Technology, Department of Biology, Cambridge, Massachusetts 02139
2 GSF-National Research Center for Environment and Health Institute of Clinical Molecular Biology and Tumor Genetics, D-81377 Munich, Germany
3 Beth Israel Deaconess Medical Center, Department of Molecular Medicine, Boston, Massachusetts 02215
4 Academic Medical Center, Department of Vascular Medicine, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands

Evans, Valerie, Antonis Hatzopoulos, William C. Aird, Helen B. Rayburn, Robert D. Rosenberg, and Jan Albert Kuivenhoven. Targeting the Hprt locus in mice reveals differential regulation of Tie2 gene expression in the endothelium. Physiol Genomics 2: 67–75, 2000.—To study the in vivo expression of the murine Tie2 gene, we have targeted the hypoxanthine phosphoribosyltransferase (Hprt) gene locus to generate two single-copy transgenic mice: T1, containing the 2,100-bp Tie2 promoter upstream from the ß-galactosidase (LacZ) gene, and T5, which also included an enhancing element originating from the first intron of the Tie2 gene. Comparing T1 and T5 embryos at day E10.5 revealed differential endothelial cell-specific expression of LacZ, whereas colocalization analyses showed that the expression was confined to endothelial cells. Moderate reporter gene activity was observed in the brain and kidney of T1 adults, whereas extensive LacZ gene expression was seen in the vasculature of most organs of the T5 adults. This study demonstrates the feasibility of targeting the Hprt locus with endothelial cell-specific sequences to analyze the spatial-temporal expression of transgenes. Of particular importance is the observation that the analysis of a single transgene copy in a defined locus allows for an accurate and rapid comparison of transcriptional activity among regulatory DNA sequences.

endothelial cells; development; transgenesis; homologous recombination; gene regulation




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