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Physiol. Genomics 2: 77-83, 2000;
1094-8341/00 $5.00
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Received 3 November 1999; accepted in final form 28 January 2000.
Physiological Genomics 2:77-83 (2000)
1094-8341/00 $5.00 © 2000 American Physiological Society

Targeting of human eNOS promoter to the Hprt locus of mice leads to tissue-restricted transgene expression

PASCALE V. GUILLOT1,*, LIXIN LIU1,*, JAN ALBERT KUIVENHOVEN2, JASON GUAN1, ROBERT D. ROSENBERG3 and WILLIAM C. AIRD1

1 Beth Israel Deaconess Medical Center, Department of Molecular Medicine, Boston 02215
2 Academic Medical Center, University of Amsterdam, Department of Vascular Medicine, 1105 AZ Amsterdam, The Netherlands
3 Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Guillot, Pascale V., Lixin Liu, Jan Albert Kuivenhoven, Jason Guan, Robert D. Rosenberg, and William C. Aird. Targeting of human eNOS promoter to the Hprt locus of mice leads to tissue-restricted transgene expression. Physiol Genomics 2: 77–83, 2000.—Phenotypic heterogeneity of the endothelium arises from cell type-specific differences in gene expression. An understanding of the mechanisms that underlie differential gene expression would provide important insight into the molecular basis of vascular diversity. In standard transgenic assays, multiple copies of heterologous DNA cassettes are randomly integrated into the mouse genome, resulting in significant line-to-line variation in expression. To overcome these limitations, we have targeted a single copy of a transgene that contains 1,600 bp of the human endothelial nitric oxide synthase (eNOS) promoter coupled to the LacZ reporter gene to the X-linked hypoxanthine phosphoribosyltransferase (Hprt) locus of mice by homologous recombination. The transgene was inserted in either of the orientations relative to that of the Hprt gene. In mice derived from multiple embryonic stem (ES) cell clones, the expression pattern was limited to a subset of endothelial cells, cardiomyocytes, and vascular smooth muscle cells. These findings suggest that Hprt locus targeting is a feasible tool for studying endothelial cell-restricted gene regulation.

gene expression; endothelial cells; transgenic mice; endothelial nitric oxide synthase; hypoxanthine phosphoribosyltransferase




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