title: Targeting gene expression to the vascular wall in transgenic mice using the murine preproendothelin-1 promoter.
abstract: To develop a system for overexpressing genes in the vascular wall, we created transgenic mice using the reporter gene luciferase and the murine preproendothelin-1 promoter. In vitro analysis suggested that the murine 5'-flanking region contained endothelial-specific elements in a 5.9-kb fragment. Five transgenic mice colonies established from independent founders all exhibited the highest level of luciferase activity in the aorta with up to 8,540 light units per microgram of protein. Immunohistochemistry with anti-luciferase antisera revealed high levels of expression in the endothelial cells of both large and small arteries and lower levels of expression in veins and capillaries. Significant expression was also seen in arterial smooth muscle cells and in select epithelial surfaces which is consistent with the known distribution of endothelin-1 in mammals. The further demonstrate the targeting capability of this system, we overexpressed the lipid-peroxidating enzyme, human 15-lipoxygenase, in the vessel wall of transgenic mice. As with luciferase, expression of active enzyme and immunohistochemical localization in vascular cells were documented in transgenic animals. Hence, this new system can be used to direct expression of molecules to the vascular wall for the purpose of examining the biological significance of either overexpression or inhibition of select proteins.
authors.affiliation: Syntex Discovery Research, Palo Alto, California 94303.
authors.forename: D
authors.initials: D
authors.lastname: Harats
authors.forename: H
authors.initials: H
authors.lastname: Kurihara
authors.forename: P
authors.initials: P
authors.lastname: Belloni
authors.forename: H
authors.initials: H
authors.lastname: Oakley
authors.forename: A
authors.initials: A
authors.lastname: Ziober
authors.forename: D
authors.initials: D
authors.lastname: Ackley
authors.forename: G
authors.initials: G
authors.lastname: Cain
authors.forename: Y
authors.initials: Y
authors.lastname: Kurihara
authors.forename: R
authors.initials: R
authors.lastname: Lawn
authors.forename: E
authors.initials: E
authors.lastname: Sigal
mesh_terms: D000818:Animals; D001011:Aorta / Q000033:anatomy & histology / Q000378:metabolism*; D001093:Arachidonate 15-Lipoxygenase / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D019332:Endothelin-1; D016232:Endothelins / Q000235:genetics*; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods*; D017930:Genes, Reporter; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000033:anatomy & histology / Q000378:metabolism; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008168:Lung / Q000033:anatomy & histology / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics*; D014018:Tissue Distribution; D014132:Trachea / Q000033:anatomy & histology / Q000378:metabolism
chemical_list: D019332:Endothelin-1; D016232:Endothelins; D011498:Protein Precursors; D001093:Arachidonate 15-Lipoxygenase; D008156:Luciferases