Endothelial-specific gene expression directed by the tie gene promoter in vivo.
The tie gene encodes a receptor tyrosine kinase that is expressed in the endothelium of blood vessels, particularly during embryonic development and angiogenesis in adults. We have cloned and characterized the mouse tie gene and isolated the human and mouse tie promoters. The promoter activities of human and mouse tie were analyzed using luciferase reporter gene constructs in transfected cell lines and beta-galactosidase constructs in transgenic mice. In transfection assays of cultured cells, both human and mouse promoter DNA fragments showed activity that was not restricted to endothelial cells. In contrast, in transgenic mice both promoters directed expression of the reporter gene to endothelial cells undergoing vasculogenesis and angiogenesis. In adult mice, tie promoter activity in lung and many vessels of the kidney was as high as in the vessels of the corresponding embryonic tissues, whereas in the heart, brain and liver, tie promoter activity was downregulated and restricted to coronaries, cusps, capillaries, and arteries. Our results show that the endothelial cell-type specificity of the tie promoter in vivo can be transferred to heterologous genes by using relatively short promoter fragments. The tie promoter, thus, has useful properties for potential gene therapy.
Korhonen
J
J
Molecular/Cancer Biology Laboratory, Haartman Institute, University of Helsinki, Finland.
Lahtinen
I
I
Halmekytö
M
M
Alhonen
L
L
Jänne
J
J
Dumont
D
D
Alitalo
K
K
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers; D004622:Embryo, Mammalian / Q000502:physiology; D005314:Embryonic and Fetal Development; D004730:Endothelium, Vascular / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation; D015698:Genomic Library; D006801:Humans; D008099:Liver / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010920:Placenta / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis* / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection
D017931:DNA Primers; D014157:Transcription Factors; D004247:DNA; D020794:Receptor Protein-Tyrosine Kinases