These SM22a-tTA/TRE-RVCH-HA bi-transgenic mice allow targeted overexpression or Tet-Off conditional expression of vascular chymase in smooth muscle cells, and may be useful in studying the role of elevated chymase activity in systemic hypertension and cardiovascular disease.
Marlene Rabinovitch, Stanford University School of Medicine
Genetic Background | Generation |
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|
Allele Type |
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Transgenic (Transactivator) |
Allele Type |
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Transgenic (Inducible, Inserted expressed sequence) |
Hemizygous tTA+/RVCH+ (or SM22a-tTA/TRE-RVCH-HA) mice are viable and fertile, harboring two transgenes using the Tet-Off system. The SM22a-tTA transgene has the 3 kb SM22alpha promoter directing expression of the tetracycline transactivator gene (tTA) to vascular smooth muscle cells (SMCs). The TRE-RVCH-HA transgene has the tetracycline-responsive element (TRE; also called tet-operator or tetO) controlling expression of a rat vascular chymase-hemagglutinin tag (RVCH-HA) fusion gene. In the absence of tetracycline (or its analog doxycycline (dox)), the SM22alpha promoter limits RVCH-HA fusion protein expression to vascular SMCs. This RVCH overexpression results in hypertension. Because this binary transgenic system also allows for a second level of control, i.e. addition of dox, expression of the RVCH-HA fusion protein can be completely abolished; reversing the hypertension phenotype. These SM22a-tTA/TRE-RVCH-HA bi-transgenic mice allow targeted overexpression or Tet-Off conditional expression of vascular chymase in smooth muscle cells, and may be useful in studying the role of elevated chymase activity in systemic hypertension and cardiovascular disease.
These mice harbor two transgenes using the Tet-Off system; SM22a-tTA and TRE-RVCH-HA. The SM22a-tTA transgene was designed with the 3 kb SM22alpha (transgelin or Tagln) promoter fragment upstream of the tetracycline transactivator gene (tTA). This construct was used to create transgenic mice on a C57BL/6 genetic background. The TRE-RVCH-HA transgene was designed with the tetracycline-responsive element (TRE; also called tet-operator or tetO) upstream of a rat vascular chymase (RVCH or Mcpt1) gene (modified such that a human influenza hemagglutinin (HA) tag is added to the C terminus of mature RVCH protein). This construct was used to create transgenic mice on an "SJL/B6" genetic background. These mice were next backcrossed to C57BL/6 mice for two to three generations and subsequently bred with the SM22a-tTA transgenic mice to generate bi-transgenic offspring (SM22a-tTA/TRE-RVCH-HA). These SM22a-tTA/TRE-RVCH-HA mice were maintained on a mixed SJL and C57BL/6 genetic background by breeding to wild-type siblings for at least 16 generations prior to arrival at The Jackson Laboratory. Upon arrival, these mice may have been bred with B6SJLF1/J (Stock No. 100012) hybrid mice to maintain the colony.
Expressed Gene | tTA, tetracycline-controlled transactivator, E. coli |
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Site of Expression | vascular smooth muscle cells (SMCs) |
Expressed Gene | Mcpt1, mast cell protease 1, rat |
Site of Expression | As the tetracycline-responsive element (TRE; also called tet-operator or tetO) controls expression of the rat vascular chymase-hemagglutinin tag (RVCH-HA) fusion gene, RVCH-HA expression is spatially and temporally dependent upon exposure to tetracycline transactivator (tTA). |
Allele Name | transgene insertion 1, Marlene Rabinovitch |
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Allele Type | Transgenic (Transactivator) |
Allele Synonym(s) | SM22alpha- tTA; tTA+ |
Gene Symbol and Name | Tg(Tagln-tTA)1Mrab, transgene insertion 1, Marlene Rabinovitch |
Gene Synonym(s) | |
Promoter | Tagln, transgelin, mouse, laboratory |
Expressed Gene | tTA, tetracycline-controlled transactivator, E. coli |
Site of Expression | vascular smooth muscle cells (SMCs) |
Strain of Origin | C57BL/6 |
Chromosome | UN |
Molecular Note | Sequence encoding a tetracycline transactivator was adjoined to the smooth muscle cell specific transgelin promoter. The promoter was reported to drive expression in the aorta, but not in the heart, lung, kidney, bladder, intestine, or stomach. |
Allele Name | transgene insertion 1, Marlene Rabinovitch |
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Allele Type | Transgenic (Inducible, Inserted expressed sequence) |
Allele Synonym(s) | RVCH+; TRE-RVCH-HA |
Gene Symbol and Name | Tg(tetO-Mcpt1)1Mrab, transgene insertion 1, Marlene Rabinovitch |
Gene Synonym(s) | |
Promoter | tetO, tet operator, |
Expressed Gene | Mcpt1, mast cell protease 1, rat |
Site of Expression | As the tetracycline-responsive element (TRE; also called tet-operator or tetO) controls expression of the rat vascular chymase-hemagglutinin tag (RVCH-HA) fusion gene, RVCH-HA expression is spatially and temporally dependent upon exposure to tetracycline transactivator (tTA). |
Strain of Origin | Not Specified |
Chromosome | UN |
Molecular Note | The transgene was designed with the tetracycline-responsive element (TRE; also called tet-operator or tetO) upstream of a rat vascular chymase (RVCH or Mcpt1) gene modified such that a human influenza hemagglutinin (HA) tag is added to the C terminus of mature RVCH protein. With addition of dox expression of the RVCH-HA fusion protein can be completely abolished, reversing the hypertension phenotype. |
When maintaining a live colony, mice hemizygous for both transgenes may be bred together, to wildtype siblings, or to B6SJLF1/J (Stock No. 100012) inbred mice. It may be useful to maintain mice on doxycycline-treated water to avoid hypertension or other incidental effects of tTA or RVCH expression.
When using the B6;SJL-Tg(Tagln-tTA)1Mrab Tg(tetO-Mcpt1)1Mrab/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #008082 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Hemizygous or non-carrier for Tg(Tagln-tTA)1Mrab ,Hemizygous or non-carrier for Tg(tetO-Mcpt1)1Mrab |
Frozen Mouse Embryo | B6;SJL-Tg(Tagln-tTA)1Mrab Tg(tetO-Mcpt1)1Mrab/J Frozen Embry | $2595.00 |
Frozen Mouse Embryo | B6;SJL-Tg(Tagln-tTA)1Mrab Tg(tetO-Mcpt1)1Mrab/J Frozen Embry | $2595.00 |
Frozen Mouse Embryo | B6;SJL-Tg(Tagln-tTA)1Mrab Tg(tetO-Mcpt1)1Mrab/J Frozen Embry | $3373.50 |
Frozen Mouse Embryo | B6;SJL-Tg(Tagln-tTA)1Mrab Tg(tetO-Mcpt1)1Mrab/J Frozen Embry | $3373.50 |
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