Replacing exons 3-4 of Ephb6 with a GFP-neo cassette reduces T cell-mediated immune responses, and affects vascular smooth muscle cell contractility and blood pressure.
Jiangping Wu, Centre Hosp. de l'Universite de Montreal
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter, Null/Knockout) | Ephb6 | Eph receptor B6 |
A GFP-neo cassette replaces exons 3-4, including the initiation sequence, of the Eph receptor B6 (Ephb6) gene, abolishing gene function. EPHB6 is a transmembrane cell surface receptor tyrosine kinase expressed on the surface of T cells, leukemic cells, vascular smooth muscle cells (VSMC), the adrenal gland medulla, skin, teeth, tongue and many brain regions. Homozygous Ephb6-/- mice are viable and fertile. Decreased EPHB6 expression occurs in mature T cells. Weak GFP fluorescence, driven by the Ephb6 promoter, is visible on the surface of CD4+CD8+ DP, CD4+ SP, and CD8+ SP thymocytes from Ephb6-/- mice. T cells from these mice show reduced expression of interleukin (IL)-2, IL-4, and interferon-γ, and have compromised responses upon activation. These mice also exhibit diminished responses to both delayed-type hypersensitivity and induced experimental autoimmune encephalitis (EAE). Small arteries from male Ephb6-/- mice display reduced contractility, while small arteries from female and castrated male Ephb6-/- mice show increased contractility. Male Ephb6-/- mice present reduced 24-h urine catechoamine excresion. Male Ephb6-/- mice present reduced 24-h urine catechoamine excresion. Levels return to normal following castration. Castrated Ephb6-/- males have increased systolic and diastolic blood pressure.
A targeting vector was designed to replace exon 3-4 of the Erythropoietin-producing hepatocyte (Eph) receptor B6 (Ephb6) gene with a green fluorescent protein (GFP)-neomycin cassette. The construct was electroporated into 129/Sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6NCrl blastocysts and the resulting chimeric mice were bred to C57BL/6NCrl mice. These mice were backcrossed to C57BL/6NCrl mice for at least ten generations. Upon arrival at The Jackson Laboratory Repository, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | Weak GFP fluorescence is visible on the surface of CD4+CD8+ DP, CD4+ SP, and CD8+ SP thymocytes. |
Allele Name | targeted mutation 1, Jiangping Wu |
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Allele Type | Targeted (Reporter, Null/Knockout) |
Allele Synonym(s) | Eph6b- |
Gene Symbol and Name | Ephb6, Eph receptor B6 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Weak GFP fluorescence is visible on the surface of CD4+CD8+ DP, CD4+ SP, and CD8+ SP thymocytes. |
Strain of Origin | 129 |
Chromosome | 6 |
Molecular Note | A sequence from the ATG in exon 3 thru the 5' end of intron 4 ws replaced with a GFP-neo cassette. Mutants demonstrated no transcript expression but did express GFP driven by the endogenous promoter. |
Mutations Made By | Jiangping Wu, Centre Hosp. de l'Universite de Montreal |
When maintaining a live colony, homozygous mice are viable and fertile.
When using the B6N.129-Ephb6tm1Jwu/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #018163 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Ephb6<tm1Jwu> |
Frozen Mouse Embryo | B6N.129-Ephb6<tm1Jwu>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6N.129-Ephb6<tm1Jwu>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6N.129-Ephb6<tm1Jwu>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6N.129-Ephb6<tm1Jwu>/J Frozen Embryo | $3373.50 |
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