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| 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. | |||||||||||
- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
- References
- Purchasing information
- Terms of Use
Description
Strain Information
Type Mutant Stock; Transgenic; Additional information on Genetically Engineered Mutant Mice. Mating System See Colony Maintenance Species laboratory mouse Generation F?+ (23-JUN-08) Donating Investigator Marlene Rabinovitch, Stanford University School of Medicine Description
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.Development
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.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 100012 B6SJLF1/J | (approximate) | |
| Considerations for Choosing Controls | ||
Related Strains
Strains carrying other alleles of Tagln
006878 B6.129S6-Taglntm2(cre)Yec/J 006875 FVB/N-Tg(Tagln-rtTA)E1Jwst/J 004746 STOCK Tg(Tagln-cre)1Her/J View Strains carrying other alleles of Tagln (3 strains)
007004 B6.Cg-Tg(Camk2a-tTA)1Mmay/DboJ 003767 B6.Cg-Tg(Eno2tTA)5021Nes/J 003763 B6.Cg-Tg(Eno2tTA)5030Nes/J 005964 B6.Cg-Tg(GFAP-tTA)110Pop/J 002618 B6.Cg-Tg(MMTVtTA)1Mam/J 006361 B6.Cg-Tg(Sp7-tTA,tetO-EGFP/cre)1Amc/J 003563 B6.Cg-Tg(tTALap)5Bjd/J 002709 B6;C3-Tg(TettTALuc)1Dgs/J 003010 B6;CBA-Tg(Camk2a-tTA)1Mmay/J 005625 FVB-Tg(Pcp2-tTA)3Horr/J 003170 FVB.Cg-Tg(Myh6-tTA)6Smbf/J 006209 FVB.Cg-Tg(Tal1-tTA)19Dgt/J 005942 FVB/N-Tg(Pf4-tTA/VP16)42Kra/J 004937 NOD.Cg-Tg(Ins2-tTA)1Doi/DoiJ 006999 STOCK Dbttm1Geh Tg(tTALap)5Bjd Tg(tetO-DBT)A1Geh/J 003272 STOCK Tg(tTALap)5Bjd/J 003271 STOCK Tg(tTAhCMV)3Bjd/J 003275 STOCK Tg(tetL)1Bjd/J 003274 STOCK Tg(tetNZL)2Bjd/J
Additional Web Information
Tet Expression Systems
Phenotype
Phenotype Information
assigned by genotype
Tg(Tagln-tTA)1Mrab/0 Tg(tetO-Mcpt1)1Mrab/0
involves: C57BL/6 * SJL
- cardiovascular system phenotype
- abnormal artery morphology (MGI Ref ID J:70031)
- medial to lumen ratio of mesenteric artery is increased by 93% compared to single transgenic controls; thickening is reversed by doxycycline-induced suppression of transgene expression
- abnormal aorta morphology (MGI Ref ID J:70031)
- increase in media to lumen ratio of aorta is observed in binary transgenic mice
- abnormal vasoconstriction (MGI Ref ID J:70031)
- endothelium-dependent (methacholine-induced) relaxation in pre-constricted arteries is impaired relative to controls
- increased vasoconstriction (MGI Ref ID J:70031)
- isolated mesenteric arteries display significantly enhanced constriction in response to phenylephrine treatment relative to control arteries
- increased left ventricle weight (MGI Ref ID J:70031)
- modest increase in left ventricle weight to body weight is observed in transgene expressing mice
- increased mean arterial blood pressure (MGI Ref ID J:70031)
- MAP is increased to 103 mm Hg compared to 82 mm Hg in controls
- increased systolic blood pressure (MGI Ref ID J:70031)
- increased by an average of 27 mm Hg relative to littermate controls; 24% increase in blood pressure in double mutants is seen compared to single transgenic mice
- doxycycline treatment to suppress transgene expression completely reverses increase, returning blood pressure to control values
- cellular phenotype
- increased cell proliferation (MGI Ref ID J:70031)
- abundant expression of proliferating cells is observed of medial layer of mesenteric artery in double mutants
This mouse can be used to support research in many areas including:
Cardiovascular Research
Hypertension (normotensive)
Hypertension
Research Tools
Cardiovascular Research (Tetop Tet System)
Genetics Research (Mutagenesis and Transgenesis: Tetop Tet System)
Tet Expression Systems (tTA/rtTA Expressing Strains)
Tet Expression Systems (tTA/rtTA Responsive Strains)
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Tg(Tagln-tTA)1Mrab | ||
|---|---|---|---|
| Allele Name | transgene insertion 1, Marlene Rabinovitch | ||
| Allele Type | Transgenic (random, expressed) | ||
| Common Name(s) | SM22alpha- tTA; tTA+; | ||
| Strain of Origin | C57BL/6 | ||
| Site of Expression | vascular smooth muscle cells (SMCs) | ||
| Expressed Gene | tTA, tetracycline-controlled transactivator, E. coli | ||
| The tetracycline-resistance gene (TetR), arose from chemically mutated Escherichia coli genome which was screened for tetracycline dependence (Gossen and Bujard, 1992). TetR was fused at the C-terminus with the viral co-activator, virion protein 16 of the herpes simplex virus (VP-16). The tetracycline-inhibitable transcription factor is a component of a bigenic system that allows doxycycline (a tetracycline analog) regulatable expression of genes that are under the direction of the tetracycline responsive promoter (TetOp)promoter. | |||
| Promoter | Tagln, transgelin, mouse, laboratory | ||
| 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. [MGI Ref ID J:70031] | ||
| Allele Symbol | Tg(tetO-Mcpt1)1Mrab | ||
| Allele Name | transgene insertion 1, Marlene Rabinovitch | ||
| Allele Type | Transgenic (random, expressed) | ||
| Common Name(s) | RVCH+; TRE-RVCH-HA; | ||
| 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). | ||
| Expressed Gene | Mcpt1, mast cell protease 1, rat | ||
| Promoter | tetO, tet operator, | ||
| 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 ofmature RVCH protein. With ddition of dox expression of the RVCH-HA fusion protein can be completely abolished, reversing the hypertension phenotype. [MGI Ref ID J:70031] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Tg(TagIn-tTA)1Mrab, STD PCR, vers. 1
Tg(tetO-Mcpt1)1Mrab, STD PCR, vers. 1
Helpful Links
Optimizing PCR Protocols
References
References
Selected Reference(s)
Ju H; Gros R; You X; Tsang S; Husain M; Rabinovitch M. 2001. Conditional and targeted overexpression of vascular chymase causes hypertension in transgenic mice. Proc Natl Acad Sci U S A 98(13):7469-74. [PubMed: 11416217] [MGI Ref ID J:70031]
Additional References
Tg(Tagln-tTA)1Mrab relatedGros R; Afroze T; You XM; Kabir G; Van Wert R; Kalair W; Hoque AE; Mungrue IN; Husain M. 2003. Plasma membrane calcium ATPase overexpression in arterial smooth muscle increases vasomotor responsiveness and blood pressure. Circ Res 93(7):614-21. [PubMed: 12933703] [MGI Ref ID J:128144]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry 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. Mating System See above Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
| Pricing for USA, Canada and Mexico shipping destinations |
|
Weeks of Age Price* Gender Genotypes Provided Individual Mouse Price $236.40 Female or Male Hemizygous for Tg(Tagln-tTA)1Mrab, Noncarrier $236.40 Female or Male Homozygous for Tg(Tagln-tTA)1Mrab, Homozygous for Tg(tetO-Mcpt1)1Mrab Individual Mouse Price $236.40 Female or Male Noncarrier, Hemizygous for Tg(Tagln-tTA)1Mrab *Price(s) in US dollars ($)
Pairs /Price* Pair Genotype $288.65 Homozygous for Tg(Tagln-tTA)1Mrab, Homozygous for Tg(tetO-Mcpt1)1Mrab x Noncarrier, Noncarrier $288.65 Noncarrier, Noncarrier x Homozygous for Tg(Tagln-tTA)1Mrab, Homozygous for Tg(tetO-Mcpt1)1Mrab
Additional Supply Details
| Supply Notes |
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|---|
| Pricing for International shipping destinations |
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Weeks of Age Price* Gender Genotypes Provided Individual Mouse Price $307.40 Female or Male Hemizygous for Tg(Tagln-tTA)1Mrab, Noncarrier $307.40 Female or Male Homozygous for Tg(Tagln-tTA)1Mrab, Homozygous for Tg(tetO-Mcpt1)1Mrab Individual Mouse Price $307.40 Female or Male Noncarrier, Hemizygous for Tg(Tagln-tTA)1Mrab *Price(s) in US dollars ($)
Pairs /Price* Pair Genotype $375.30 Homozygous for Tg(Tagln-tTA)1Mrab, Homozygous for Tg(tetO-Mcpt1)1Mrab x Noncarrier, Noncarrier $375.30 Noncarrier, Noncarrier x Homozygous for Tg(Tagln-tTA)1Mrab, Homozygous for Tg(tetO-Mcpt1)1Mrab
Additional Supply Details
| Supply Notes |
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Supply Details
| Standard Supply | Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of ~nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within 48 hours of order placement. |
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| Supply Notes |
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Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 100012 B6SJLF1/J | (approximate) | |
| Considerations for Choosing Controls | ||
| USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
| International - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
General Terms and Conditions
See Terms of Use
The Jackson Laboratory's Genotype Promise
The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.Ordering and Purchasing Information
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Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
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Terms of Use
Terms of Use
General Terms and Conditions
Use of the Tet-System may require a license, see Licenses for Strains Using TET-System Technology.
For additional Licensing and Use Restrictions view the link(s) below:
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Contact information
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| phone: | 207-288-6470 |
| fax: | 207-288-6655 |
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