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- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
- References
- Purchasing information
- Terms of Use
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) 14-APR-08 Species laboratory mouse Generation N5F5 (18-NOV-09) Donating Investigator IMR Colony, The Jackson Laboratory Description
Mice that are hemizygous for this transgenic insert are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. These transgenic mice express the reverse tetracycline-controlled transactivator (rtTA) protein under the control of the human SFTPC, surfactant, pulmonary-associated protein C, promoter. In situ hybridization detects rtTA gene product (mRNA) in lung peripheral epithelial cells from adult mice and 15 postconception day aged embryos from doxycycline treated dams. Induction of transgene expression is detected as early as postconception day 12.5 when the pregnant female is treated with doxycycline. When mated to a second transgenic strain carrying a gene of interest under the regulatory control of a tetracycline-responsive promoter element (tetO), expression of the target gene may be regulated by the tetracycline analog, doxycycline (dox); in the presence of dox, transcription of the target gene is induced in cells where rtTA is expressed. This strain provides a Tet-On tool that allows the inducible expression of genes in the developing and adult lung and respiratory epithelium.In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. This is the case for this strain. 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
A transgenic construct containing 3.7kb sequence of the human SFTPC, surfactant, pulmonary-associated protein C, gene promoter, the reverse tetracycline regulatable transactivator protein (rtTA) gene, and SV40 polyadenylation signal sequence was injected into mixed C57BL/6 and 129 background embryos. Founder mice were bred to FVB/N animals. Upon arrival at The Jackson Laboratory, these mice were backcrossed to C57BL/6J mice for at least 5 generations.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
Related Strains
Strains carrying Tg(SFTPC-rtTA)5Jaw allele
006245 C.Cg-Tg(SFTPC-rtTA)5Jaw/J 006225 FVB.Cg-Tg(SFTPC-rtTA)5Jaw/J View Strains carrying Tg(SFTPC-rtTA)5Jaw (2 strains)
Additional Web Information
Tet Expression Systems
Phenotype
Phenotype Information
This mouse can be used to support research in many areas including:
Developmental Biology Research
Internal/Organ Defects
lung
Immunology and Inflammation Research
Cystic Fibrosis
Internal/Organ Research
Lung Defects
Mouse/Human Gene Homologs
cystic fibrosis
Research Tools
Cancer Research
Tetop Tet System
Developmental Biology Research
Genetics Research
Mutagenesis and Transgenesis: Tetop Tet System
Immunology and Inflammation Research
Internal/Organ Research
Tet Expression Systems
tTA/rtTA Expressing Strains
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Tg(SFTPC-rtTA)5Jaw | ||
|---|---|---|---|
| Allele Name | transgene insertion 5, Jeffrey A Whitsett | ||
| Allele Type | Transgenic (random, expressed) | ||
| Common Name(s) | SP-C-rtTAtg; SP-CrtTA; SP-CrtTAtg; SPC-rtTA; Sp-c-rtta; Tg(SFTPC-rtTA)1Jaw; hSPC-rtTA+; | ||
| Mutation Made By | Jeffrey Whitsett, Children's Hospital Medical Center | ||
| Site of Expression | Expresses rtTA in developing and adult lung. | ||
| Expressed Gene | rtTA, reverse tetracycline-controlled transactivator, E. coli | ||
| The tetracycline repressor gene (Tetr), arose from chemically mutated Escherichia coli genome which was screened for tetracycline dependence (Gossen and Bujard, 1992). One mutant with a four amino acid residue change (rTetR) exhibited dependence on tetracycline for induction of the targeted gene and was used in the rtTA construct (Gossen et al, 1995). rTetr was fused at the C-terminus with the viral co-activator, virion protein 16 of the herpes simplex virus (VP-16). | |||
| Promoter | SFTPC, surfactant protein C, human | ||
| Molecular Note | The transgene is composed of 3.7 kb of human SFTPC promoter, a reverse tetracycline responsive transactivator gene (rt-TA), and a 0.45 kb SV40 polyadenylation signal. Transgene expression was confirmed by RT-PCR analysis. The human SFTPC promoter is active in respiratory epithelial cells. [MGI Ref ID J:61736] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Tg(tTA), Melt Curve Analysis
Tg(tTA), QPCR
Tg(tTA), Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
Tichelaar JW; Lu W; Whitsett JA. 2000. Conditional expression of fibroblast growth factor-7 in the developing and mature lung. J Biol Chem 275(16):11858-64. [PubMed: 10766812] [MGI Ref ID J:61736]
Additional References
Tg(SFTPC-rtTA)5Jaw relatedAkeson AL; Greenberg JM; Cameron JE; Thompson FY; Brooks SK; Wiginton D; Whitsett JA. 2003. Temporal and spatial regulation of VEGF-A controls vascular patterning in the embryonic lung. Dev Biol 264(2):443-55. [PubMed: 14651929] [MGI Ref ID J:86840]
Basseres DS; Levantini E; Ji H; Monti S; Elf S; Dayaram T; Fenyus M; Kocher O; Golub T; Wong KK; Halmos B; Tenen DG. 2006. Respiratory failure due to differentiation arrest and expansion of alveolar cells following lung-specific loss of the transcription factor C/EBPalpha in mice. Mol Cell Biol 26(3):1109-23. [PubMed: 16428462] [MGI Ref ID J:105593]
Ceteci F; Ceteci S; Karreman C; Kramer BW; Asan E; Gotz R; Rapp UR. 2007. Disruption of Tumor Cell Adhesion Promotes Angiogenic Switch and Progression to Micrometastasis in RAF-Driven Murine Lung Cancer. Cancer Cell 12(2):145-159. [PubMed: 17692806] [MGI Ref ID J:124321]
Chen H; Zhuang F; Liu YH; Xu B; Del Moral P; Deng W; Chai Y; Kolb M; Gauldie J; Warburton D; Moses HL; Shi W. 2008. TGF-beta receptor II in epithelia versus mesenchyme plays distinct roles in the developing lung. Eur Respir J 32(2):285-95. [PubMed: 18321928] [MGI Ref ID J:137996]
Clark JC; Tichelaar JW; Wert SE; Itoh N; Perl AK; Stahlman MT; Whitsett JA. 2001. FGF-10 disrupts lung morphogenesis and causes pulmonary adenomas in vivo. Am J Physiol Lung Cell Mol Physiol 280(4):L705-15. [PubMed: 11238011] [MGI Ref ID J:68525]
Dave V; Childs T; Xu Y; Ikegami M; Besnard V; Maeda Y; Wert SE; Neilson JR; Crabtree GR; Whitsett JA. 2006. Calcineurin/Nfat signaling is required for perinatal lung maturation and function. J Clin Invest 116(10):2597-609. [PubMed: 16998587] [MGI Ref ID J:114501]
Dave V; Wert SE; Tanner T; Thitoff AR; Loudy DE; Whitsett JA. 2008. Conditional deletion of Pten causes bronchiolar hyperplasia. Am J Respir Cell Mol Biol 38(3):337-45. [PubMed: 17921358] [MGI Ref ID J:146355]
Floyd HS; Farnsworth CL; Kock ND; Mizesko MC; Little JL; Dance ST; Everitt J; Tichelaar J; Whitsett JA; Miller MS. 2005. Conditional expression of the mutant Ki-rasG12C allele results in formation of benign lung adenomas: development of a novel mouse lung tumor model. Carcinogenesis 26(12):2196-206. [PubMed: 16051643] [MGI Ref ID J:102839]
Gontan C; de Munck A; Vermeij M; Grosveld F; Tibboel D; Rottier R. 2008. Sox2 is important for two crucial processes in lung development: branching morphogenesis and epithelial cell differentiation. Dev Biol 317(1):296-309. [PubMed: 18374910] [MGI Ref ID J:136180]
Hokuto I; Ikegami M; Yoshida M; Takeda K; Akira S; Perl AK; Hull WM; Wert SE; Whitsett JA. 2004. Stat-3 is required for pulmonary homeostasis during hyperoxia. J Clin Invest 113(1):28-37. [PubMed: 14702106] [MGI Ref ID J:87622]
Kalin TV; Wang IC; Meliton L; Zhang Y; Wert SE; Ren X; Snyder J; Bell SM; Graf L Jr; Whitsett JA; Kalinichenko VV. 2008. Forkhead Box m1 transcription factor is required for perinatal lung function. Proc Natl Acad Sci U S A 105(49):19330-5. [PubMed: 19033457] [MGI Ref ID J:142246]
Kida H; Mucenski ML; Thitoff AR; Le Cras TD; Park KS; Ikegami M; Muller W; Whitsett JA. 2008. GP130-STAT3 regulates epithelial cell migration and is required for repair of the bronchiolar epithelium. Am J Pathol 172(6):1542-54. [PubMed: 18467707] [MGI Ref ID J:136187]
Kim KK; Wei Y; Szekeres C; Kugler MC; Wolters PJ; Hill ML; Frank JA; Brumwell AN; Wheeler SE; Kreidberg JA; Chapman HA. 2009. Epithelial cell alpha3beta1 integrin links beta-catenin and Smad signaling to promote myofibroblast formation and pulmonary fibrosis. J Clin Invest 119(1):213-24. [PubMed: 19104148] [MGI Ref ID J:144703]
Kulkarni RM; Greenberg JM; Akeson AL. 2009. NFATc1 regulates lymphatic endothelial development. Mech Dev 126(5-6):350-65. [PubMed: 19233265] [MGI Ref ID J:149253]
Lin S; Ikegami M; Xu Y; Bosserhoff AK; Malkinson AM; Shannon JM. 2008. Misexpression of MIA disrupts lung morphogenesis and causes neonatal death. Dev Biol 316(2):441-55. [PubMed: 18342301] [MGI Ref ID J:135990]
Mallory BP; Mead TJ; Wiginton DA; Kulkarni RM; Greenberg JM; Akeson AL. 2006. Lymphangiogenesis in the developing lung promoted by VEGF-A. Microvasc Res 72(1-2):62-73. [PubMed: 16806288] [MGI Ref ID J:129289]
Martis PC; Whitsett JA; Xu Y; Perl AK; Wan H; Ikegami M. 2006. C/EBP{alpha} is required for lung maturation at birth. Development 133(6):1155-64. [PubMed: 16467360] [MGI Ref ID J:106543]
Matsuzaki Y; Besnard V; Clark JC; Xu Y; Wert SE; Ikegami M; Whitsett JA. 2008. STAT3 regulates ABCA3 expression and influences lamellar body formation in alveolar type II cells. Am J Respir Cell Mol Biol 38(5):551-8. [PubMed: 18096869] [MGI Ref ID J:149721]
Matsuzaki Y; Xu Y; Ikegami M; Besnard V; Park KS; Hull WM; Wert SE; Whitsett JA. 2006. Stat3 is required for cytoprotection of the respiratory epithelium during adenoviral infection. J Immunol 177(1):527-37. [PubMed: 16785550] [MGI Ref ID J:134441]
Metzger DE; Stahlman MT; Shannon JM. 2008. Misexpression of ELF5 disrupts lung branching and inhibits epithelial differentiation. Dev Biol 320(1):149-60. [PubMed: 18544451] [MGI Ref ID J:139210]
Miller LA; Wert SE; Clark JC; Xu Y; Perl AK; Whitsett JA. 2004. Role of Sonic hedgehog in patterning of tracheal-bronchial cartilage and the peripheral lung. Dev Dyn 231(1):57-71. [PubMed: 15305287] [MGI Ref ID J:91723]
Morimoto M; Kopan R. 2009. rtTA toxicity limits the usefulness of the SP-C-rtTA transgenic mouse. Dev Biol 325(1):171-8. [PubMed: 19013447] [MGI Ref ID J:143537]
Mucenski ML; Wert SE; Nation JM; Loudy DE; Huelsken J; Birchmeier W; Morrisey EE; Whitsett JA. 2003. beta-Catenin is required for specification of proximal/distal cell fate during lung morphogenesis. J Biol Chem 278(41):40231-8. [PubMed: 12885771] [MGI Ref ID J:85946]
Nguyen NM; Kelley DG; Schlueter JA; Meyer MJ; Senior RM; Miner JH. 2005. Epithelial laminin alpha5 is necessary for distal epithelial cell maturation, VEGF production, and alveolization in the developing murine lung. Dev Biol 282(1):111-25. [PubMed: 15936333] [MGI Ref ID J:104565]
Park KS; Wells JM; Zorn AM; Wert SE; Laubach VE; Fernandez LG; Whitsett JA. 2006. Transdifferentiation of ciliated cells during repair of the respiratory epithelium. Am J Respir Cell Mol Biol 34(2):151-7. [PubMed: 16239640] [MGI Ref ID J:120191]
Park KS; Wells JM; Zorn AM; Wert SE; Whitsett JA. 2006. Sox17 influences the differentiation of respiratory epithelial cells. Dev Biol 294(1):192-202. [PubMed: 16574095] [MGI Ref ID J:109303]
Perl AK; Gale E. 2009. FGF signaling is required for myofibroblast differentiation during alveolar regeneration. Am J Physiol Lung Cell Mol Physiol 297(2):L299-308. [PubMed: 19502291] [MGI Ref ID J:151422]
Perl AK; Kist R; Shan Z; Scherer G; Whitsett JA. 2005. Normal lung development and function after Sox9 inactivation in the respiratory epithelium. Genesis 41(1):23-32. [PubMed: 15645446] [MGI Ref ID J:95910]
Perl AK; Tichelaar JW; Whitsett JA. 2002. Conditional gene expression in the respiratory epithelium of the mouse. Transgenic Res 11(1):21-9. [PubMed: 11874100] [MGI Ref ID J:129109]
Perl AK; Wert SE; Loudy DE; Shan Z; Blair PA; Whitsett JA. 2005. Conditional recombination reveals distinct subsets of epithelial cells in trachea, bronchi, and alveoli. Am J Respir Cell Mol Biol 33(5):455-62. [PubMed: 16055670] [MGI Ref ID J:132772]
Perl AK; Wert SE; Nagy A; Lobe CG; Whitsett JA. 2002. Early restriction of peripheral and proximal cell lineages during formation of the lung. Proc Natl Acad Sci U S A 99(16):10482-7. [PubMed: 12145322] [MGI Ref ID J:78365]
Quinton LJ; Jones MR; Robson BE; Simms BT; Whitsett JA; Mizgerd JP. 2008. Alveolar epithelial STAT3, IL-6 family cytokines, and host defense during Escherichia coli pneumonia. Am J Respir Cell Mol Biol 38(6):699-706. [PubMed: 18192501] [MGI Ref ID J:149718]
Rapp UR; Korn C; Ceteci F; Karreman C; Luetkenhaus K; Serafin V; Zanucco E; Castro I; Potapenko T. 2009. MYC is a metastasis gene for non-small-cell lung cancer. PLoS One 4(6):e6029. [PubMed: 19551151] [MGI Ref ID J:150188]
Romano RA; Ortt K; Birkaya B; Smalley K; Sinha S. 2009. An active role of the DeltaN isoform of p63 in regulating basal keratin genes K5 and K14 and directing epidermal cell fate. PLoS One 4(5):e5623. [PubMed: 19461998] [MGI Ref ID J:149319]
Saini Y; Harkema JR; LaPres JJ. 2008. HIF1alpha is essential for normal intrauterine differentiation of alveolar epithelium and surfactant production in the newborn lung of mice. J Biol Chem 283(48):33650-7. [PubMed: 18801745] [MGI Ref ID J:143384]
Shaw AT; Meissner A; Dowdle JA; Crowley D; Magendantz M; Ouyang C; Parisi T; Rajagopal J; Blank LJ; Bronson RT; Stone JR; Tuveson DA; Jaenisch R; Jacks T. 2007. Sprouty-2 regulates oncogenic K-ras in lung development and tumorigenesis. Genes Dev 21(6):694-707. [PubMed: 17369402] [MGI Ref ID J:119477]
Shu W; Guttentag S; Wang Z; Andl T; Ballard P; Lu MM; Piccolo S; Birchmeier W; Whitsett JA; Millar SE; Morrisey EE. 2005. Wnt/beta-catenin signaling acts upstream of N-myc, BMP4, and FGF signaling to regulate proximal-distal patterning in the lung. Dev Biol 283(1):226-39. [PubMed: 15907834] [MGI Ref ID J:99391]
Sun J; Chen H; Chen C; Whitsett JA; Mishina Y; Bringas P Jr; Ma JC; Warburton D; Shi W. 2008. Prenatal lung epithelial cell-specific abrogation of alk3-bone morphogenetic protein signaling causes neonatal respiratory distress by disrupting distal airway formation. Am J Pathol 172(3):571-82. [PubMed: 18258849] [MGI Ref ID J:132011]
Tian Y; Zhou R; Rehg JE; Jackowski S. 2007. Role of phosphocholine cytidylyltransferase alpha in lung development. Mol Cell Biol 27(3):975-82. [PubMed: 17130238] [MGI Ref ID J:118292]
Wan H; Dingle S; Xu Y; Besnard V; Kaestner KH; Ang SL; Wert S; Stahlman MT; Whitsett JA. 2005. Compensatory roles of Foxa1 and Foxa2 during lung morphogenesis. J Biol Chem 280(14):13809-16. [PubMed: 15668254] [MGI Ref ID J:98750]
Wan H; Kaestner KH; Ang SL; Ikegami M; Finkelman FD; Stahlman MT; Fulkerson PC; Rothenberg ME; Whitsett JA. 2004. Foxa2 regulates alveolarization and goblet cell hyperplasia. Development 131(4):953-64. [PubMed: 14757645] [MGI Ref ID J:88601]
Wan H; Luo F; Wert SE; Zhang L; Xu Y; Ikegami M; Maeda Y; Bell SM; Whitsett JA. 2008. Kruppel-like factor 5 is required for perinatal lung morphogenesis and function. Development 135(15):2563-72. [PubMed: 18599506] [MGI Ref ID J:138576]
Wan H; Xu Y; Ikegami M; Stahlman MT; Kaestner KH; Ang SL; Whitsett JA. 2004. Foxa2 is required for transition to air breathing at birth. Proc Natl Acad Sci U S A 101(40):14449-54. [PubMed: 15452354] [MGI Ref ID J:93473]
Wang IC; Meliton L; Ren X; Zhang Y; Balli D; Snyder J; Whitsett JA; Kalinichenko VV; Kalin TV. 2009. Deletion of Forkhead Box M1 transcription factor from respiratory epithelial cells inhibits pulmonary tumorigenesis. PLoS One 4(8):e6609. [PubMed: 19672312] [MGI Ref ID J:152467]
White AC; Lavine KJ; Ornitz DM. 2007. FGF9 and SHH regulate mesenchymal Vegfa expression and development of the pulmonary capillary network. Development 134(20):3743-52. [PubMed: 17881491] [MGI Ref ID J:128368]
Xu J; Tian J; Grumelli SM; Haley KJ; Shapiro SD. 2006. Stage-specific effects of cAMP signaling during distal lung epithelial development. J Biol Chem 281(50):38894-904. [PubMed: 17018522] [MGI Ref ID J:117640]
Yamamoto H; Yun EJ; Gerber HP; Ferrara N; Whitsett JA; Vu TH. 2007. Epithelial-vascular cross talk mediated by VEGF-A and HGF signaling directs primary septae formation during distal lung morphogenesis. Dev Biol 308(1):44-53. [PubMed: 17583691] [MGI Ref ID J:124125]
Yanagi S; Kishimoto H; Kawahara K; Sasaki T; Sasaki M; Nishio M; Yajima N; Hamada K; Horie Y; Kubo H; Whitsett JA; Mak TW; Nakano T; Nakazato M; Suzuki A. 2007. Pten controls lung morphogenesis, bronchioalveolar stem cells, and onset of lung adenocarcinomas in mice. J Clin Invest 117(10):2929-40. [PubMed: 17909629] [MGI Ref ID J:127405]
Yang YS; Yang MC; Guo Y; Williams OW; Weissler JC. 2009. PLAGL2 expression-induced lung epithelium damages at bronchiolar alveolar duct junction in emphysema: bNip3- and SP-C-associated cell death/injury activity. Am J Physiol Lung Cell Mol Physiol 297(3):L455-66. [PubMed: 19574421] [MGI Ref ID J:152422]
Yin Y; White AC; Huh SH; Hilton MJ; Kanazawa H; Long F; Ornitz DM. 2008. An FGF-WNT gene regulatory network controls lung mesenchyme development. Dev Biol 319(2):426-36. [PubMed: 18533146] [MGI Ref ID J:137691]
Zhang Y; Goss AM; Cohen ED; Kadzik R; Lepore JJ; Muthukumaraswamy K; Yang J; DeMayo FJ; Whitsett JA; Parmacek MS; Morrisey EE. 2008. A Gata6-Wnt pathway required for epithelial stem cell development and airway regeneration. Nat Genet 40(7):862-70. [PubMed: 18536717] [MGI Ref ID J:138407]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, these mice are bred as hemizygotes. Mating System Noncarrier x Hemizygote (Female x Male) 14-APR-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 |
|
Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $243.50 Female or Male Hemizygous for Tg(SFTPC-rtTA)5Jaw
Pairs /Price (US dollars $) Pair Genotype $297.85 Hemizygous for Tg(SFTPC-rtTA)5Jaw x Noncarrier $297.85 Noncarrier x Hemizygous for Tg(SFTPC-rtTA)5Jaw
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(SFTPC-rtTA)5Jaw
Pairs /Price (US dollars $) Pair Genotype $387.30 Hemizygous for Tg(SFTPC-rtTA)5Jaw x Noncarrier $387.30 Noncarrier x Hemizygous for Tg(SFTPC-rtTA)5Jaw
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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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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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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JAX® Mice, Products & Services Conditions of Use
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