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| These smMHC/Cre/eGFP transgenic mice may be useful in studies utilizing "Cre-lox" technology or fluorescent protein expression in smooth muscle, especially separation of vascular myocytes from the surrounding cellular environment to isolate muscle specific adaptations or disease responses. | |||||||||||||||
- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
- References
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Description
Strain Information
Type Congenic; Mutant Strain; Transgenic; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Additional information on Congenic nomenclature. Mating System Noncarrier x Hemizygote (Female x Male) 02-JUL-08 Species laboratory mouse Generation N13+N1F1 (18-DEC-08) Donating Investigator Michael Kotlikoff, Cornell University Description
Mice hemizygous for the smMHC/Cre/eGFP transgene (smMHCCre/eGFP) are viable and fertile, with the smooth muscle myosin heavy chain (smMHC or Myh11) promoter directing bicistronic Cre and EGFP protein expression to smooth muscle cells during development as well as in the adult mouse. Hemizygotes from founder line SMCG2 (SM2Cre/GFP) display intense EGFP fluorescence restricted to vascular and nonvascular smooth muscle, with strong concordance between cre expression and EGFP fluorescence (verifying the use of fluorescence as a marker for conditional gene recombination). When bred with mice containing a loxP-flanked sequence of interest, the resulting offspring can have Cre-mediated recombination of the flanked sequence in smooth muscle. Homozygotes are viable and fertile, with smaller litter sizes and a higher incidence of perinatal mortality. These smMHC/Cre/eGFP transgenic mice may be useful in studies utilizing "Cre-lox" technology or fluorescent protein expression in smooth muscle, especially separation of vascular myocytes from the surrounding cellular environment to isolate muscle specific adaptations or disease responses.Considerations for experimental use: The donating investigator reports that the transgene is expressed in female germ cells and that Cre protein (or maternal RNA) may be passed to all embryos. If bred to males carrying a "floxed" allele, all offspring (cre-positive or cre-negative) may exhibit universal Cre-mediated recombination of the allele during early embryonic development. The donating investigator also reports transgene expression in the male germ line; post meiotic spermatagonia transiently express Cre recombinase with expression ending prior to fertilization. Therefore, Cre activity may be observed in all haploid male germ cells (cre-positive or cre-negative). If transgenic males also harbor "floxed" alleles in their genome, these loci may undergo universal Cre-mediated recombination in the sperm. As Cre activity ceases before fertilization, the resulting offspring exhibit no universal deletion affect on any "floxed" allele(s) of maternal origin. Because smMHC/Cre/eGFP transgene expression is observed in both maternal and paternal germlines, it is recommended to maintain the smMHC/Cre/EGFP transgenic colony without additional "floxed" mutations in their genome.
Development
The smMHC/Cre/eGFP transgene was designed placing a Cre recombinase gene, internal ribosomal entry site (IRES), and an enhanced green fluorescent protein (EGFP) gene all downstream of the 16 kb mouse smooth muscle myosin heavy chain (smMHC or Myh11) promoter fragment. This transgene was microinjected into the pronucleus of B6D2F2 fertilized mouse eggs. Transgenic mice were bred to C57BL/6J, and the resulting transgenic offspring with a single integration site and highest copy number (founder line SMCG2) were established. These smMHC/Cre/eGFP mice were backcrossed to C57BL/6J inbred mice for at least 12 generations prior to arrival at The Jackson Laboratory. During the backcross, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
View Fluorescent Protein Strains (225 strains)
Additional Web Information
Fluorescent Proteins/lacZ Systems
Introduction to Cre-lox technology
Phenotype
Phenotype Information
This mouse can be used to support research in many areas including:
GFP relatedCardiovascular Research
Other
Developmental Biology Research
Perinatal Lethality
Homozygous
Research Tools
Cardiovascular Research
Cre-lox System
Cre-lox System
Cre Recombinase Expression
Cre Recombinase Expression: Germline/Embryonic Expression
Developmental Biology Research
Cre-lox System
transplantation marker for embryonic and adult tissue
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers: Cre-lox System
Tissue/Cell Markers: multiple
Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
Neurobiology Research
cell marker
cre relatedResearch Tools
Fluorescent Proteins
Cre-lox System
Genetics Research
Mutagenesis and Transgenesis: Cre-lox System
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Tg(Myh11-cre,-EGFP)2Mik | ||
|---|---|---|---|
| Allele Name | transgene insertion 2, M I Kotlikoff | ||
| Allele Type | Transgenic (Cre/Flp) | ||
| Common Name(s) | Tg(Myh11-cre/EGFP)1Mik; smMHC-Cre; smMHCCre/eGFP; | ||
| Mutation Made By | Michael Kotlikoff, Cornell University | ||
| Strain of Origin | (C57BL/6 x DBA/2)F2 | ||
| Site of Expression | EGFP and Cre recombinase are expressed in vascular and nonvascular smooth muscle during development and in the adult mouse. | ||
| Expressed Gene | cre, cre recombinase, bacteriophage P1 | ||
| Cre recombinase is an enzyme derived from the bacteriophage P1 that specifically recognizes loxP sites. Cre has been shown to effectively mediate the excision of DNA located between loxP sites. After the excision event, the DNA ends recombine leaving a single loxP site in place of the intervening sequence. | |||
| Expressed Gene | GFP, Green Fluorescent Protein, jellyfish | ||
| Green Fluorescent Protein (GFP), derived from the jellyfish Aequorea victoria, is a versatile reporter molecule which has found use in many biological applications. In some constructs the original molecule has been modified in order to enhance its fluorescence intensity (EGFP, enhanced GFP). When utilized in a transgenic construct, tissue expressing sufficient amounts of GFP will fluoresce when exposed to a 488 nm light source. | |||
| Promoter | Myh11, myosin, heavy polypeptide 11, smooth muscle, mouse, laboratory | ||
| Driver Note | Myh11 | ||
| General Note | Two transgenic founders, SMCG2 and SMCG3, were created. Transgenic founders were bred to C57BL/6 mice. | ||
| Molecular Note | The transgene consists of a mouse Myh11 promoter, a cre recombinase gene, an internal ribosome entry site (IRES) and an enhanced GFP gene. 16 kb of mouse Myh11 promoter sequences was used to drive the bicistronic transgene expression in smooth muscle cells. [MGI Ref ID J:81358] | ||
| Gene Symbol and Name | Tg(Myh11-cre,-EGFP)2Mik, transgene insertion 2, M I Kotlikoff | ||
| Chromosome | UN | ||
| Gene Common Name(s) | Tg(Myh11-cre/EGFP)1Mik; smMHC-Cre; smMHCCre/eGFP; transgene insertion 1, M I Kotlikoff; | ||
Genotyping
Genotyping Information
Genotyping Protocols
Fluorescent Proteins (Generic GFP), Standard PCR
Generic Cre Melt Curve Analysis, Melt Curve Analysis
Generic Cre, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
Xin HB; Deng KY; Rishniw M; Ji G; Kotlikoff MI. 2002. Smooth muscle expression of Cre recombinase and eGFP in transgenic mice. Physiol Genomics 10(3):211-5. [PubMed: 12209023] [MGI Ref ID J:81358]
Additional References
Tg(Myh11-cre,-EGFP)2Mik relatedCohn HI; Harris DM; Pesant S; Pfeiffer M; Zhou RH; Koch WJ; Dorn GW nd; Eckhart AD. 2008. Inhibition of vascular smooth muscle G protein-coupled receptor kinase 2 enhances alpha1D-adrenergic receptor constriction. Am J Physiol Heart Circ Physiol 295(4):H1695-704. [PubMed: 18723764] [MGI Ref ID J:141312]
Roh M; Kim J; Song C; Wills M; Abdulkadir SA. 2006. Transgenic mice for Cre-inducible overexpression of the oncogenes c-MYC and Pim-1 in multiple tissues. Genesis 44(10):447-53. [PubMed: 17013838] [MGI Ref ID J:114516]
de Lange WJ; Halabi CM; Beyer AM; Sigmund CD. 2008. Germ Line Activation of the Tie2 and SMMHC Promoters Causes Non-Cell Specific Deletion of Floxed Alleles. Physiol Genomics :. [PubMed: 18612081] [MGI Ref ID J:138838]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, hemizygous smMHC/Cre/EGFP mice can be bred to wild-type siblings or C57BL/6J inbred mice. Homozygotes are viable and fertile, with smaller litter sizes and a higher incidence of perinatal mortality. Because transgene expression is observed in both maternal and paternal germlines (see Strain Phenotype description for details), it is recommended to maintain the smMHC/Cre/EGFP transgenic colony without additional "floxed" mutations in their genome. Mating System Noncarrier x Hemizygote (Female x Male) 02-JUL-08 Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
| Pricing for USA, Canada and Mexico shipping destinations |
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Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $243.50 Female or Male Hemizygous for Tg(Myh11-cre,-EGFP)2Mik
Pairs /Price (US dollars $) Pair Genotype $297.85 Hemizygous for Tg(Myh11-cre,-EGFP)2Mik x Noncarrier $297.85 Noncarrier x Hemizygous for Tg(Myh11-cre,-EGFP)2Mik
Additional Supply Details
| Pricing for International shipping destinations |
|
Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $316.60 Female or Male Hemizygous for Tg(Myh11-cre,-EGFP)2Mik
Pairs /Price (US dollars $) Pair Genotype $387.30 Hemizygous for Tg(Myh11-cre,-EGFP)2Mik x Noncarrier $387.30 Noncarrier x Hemizygous for Tg(Myh11-cre,-EGFP)2Mik
Additional Supply Details
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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 approximately 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 two business days following order placement. |
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| Supply Notes |
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Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 000664 C57BL/6J | ||
| 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. | ||
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