STOCK Tg(Tek-rtTA,tetO-lacZ)1425Tpr/J

Stock number: 005493
Product name: Tie2-rTA/TRE-βGal
Research areas: Research Tools

Description

These transgenic mice harbor the reverse tetracycline-controlled transactivator (rtTA) and the beta-galactosidase gene (lacZ), and in the presence of tetracycline or a tetracycline analog, expression of lacZ is observed in the nuclei of vascular endothelium in a wide variety of tissues.

Detailed description

Hemizygous transgenic mice are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. These transgenic mice harbor co-injected transgenic constructs. The reverse tetracycline-controlled transactivator (rtTA) protein is expressed under the direction of the endothelial-specific receptor tyrosine kinase enhancer/promoter (Tek). The second transgenic construct expresses a nuclear-localizing beta-galactosidase gene (lacZ) under the control of a tetracycline-responsive element (TRE). In the presence of tetracycline or a tetracycline analog (such as doxycycline), selective expression of lacZ is observed in the nuclei of vascular endothelium in a wide variety of tissues (aorta, heart, brain, lung, kidney, liver, spleen, uterus, prostate, stomach, skeletal muscle, large and small intestine). Expression is observed as early as embryonic day 9.5. In the absence of tetracycline, some lacZ expression occurs in the smaller branches of the aorta. Additionally, the donating investigators report a trend towards diminished lacZ expression when the alleles are crossed onto a C57BL/6 background. This strain provides a tool that allows the inducible expression of genes in the vascular endothelium during early developmental stages and in the adult.

This strain is on a genetic background different from that on which the allele was first characterized. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.

Development

To generate this transgenic line, two transgenic constructs were co-injected into B6CBA fertilized oocytes. The first construct included a 2.1 kb endothelial-specific receptor tyrosine kinase (Tek) promoter fragment, the Tet-on transactivator (tTA), SV40 polyadenylation sequence and a 1.7 kb enhancer sequence derived from Tek intron 1. The other construct consisted of a tetracycline-responsive element, nuclear-localizing beta-galactosidase (lacZ) gene and a SV40 polyadenylation sequence. Although the transgenic constructs are closely linked in this transgenic line (1425), the donating investigators report that no definitive evidence exits indicating that they have cointegrated. The mice were later crossed to Apoe mutant mice (that may have been congenic on the C57BL/6 background). During the development of the colony at The Jackson Laboratory, mice were selected for the (Tek-rtTA,TRE-lacZ) transgene and against the Apoe mutant allele.

Allele

Symbol: Tg(Tek-rtTA,tetO-lacZ)1425Tpr
Name: transgene insertion 1425, James N Topper
MGI accession ID: MGI:3057127
Generation method: Transgenic
Attributes: Reporter; Inducible; Transactivator
Marker symbol: Tg(Tek-rtTA,tetO-lacZ)1425Tpr
Marker name: transgene insertion 1425, James N Topper
Chromosome: UN
Strain of origin: (C57BL/6 x CBA)F1
Expressed genes: rtTA,reverse tetracycline-controlled transactivator,E. coli; lacZ,beta-galactosidase,E. coli
Site of expression: Expresses rtTA in vascular endothelial cells.
Molecular note: Two transgenic constructs were co-injected into B6CBA fertilized oocytes. The first construct included a 2.1 kb endothelial-specific receptor tyrosine kinase (Tek) promoter fragment, the Tet-on transactivator, SV40 polyadenylation sequence, and a 1.7 kb enhancer sequence derived from Tek intron 1. The other construct consisted of a tetracycline-responsive element, nuclear-localizing beta-galactosidase gene, and a SV40 polyadenylation sequence. Although the transgenic constructs are closely linked in transgenic line 1425, no definitive evidence exists indicating that they have cointegrated.
General note: Hemizygous transgenic mice are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. In the presence of tetracycline or a tetracycline analog such as doxycycline, selective expression of beta-galactosidase is observed in the nuclei of vascular endothelium in a wide variety of tissues: aorta, heart, brain, lung, kidney, liver, spleen, uterus, prostate, stomach, skeletal muscle, large and small intestine. Expression is observed as early as embryonic day 9.5. In the absence of tetracycline, some beta-galactosidase expression occurs in the smaller branches of the aorta. A trend towards diminished beta-galactosidase expression when the alleles are crossed onto a C57BL/6 background has been reported.