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Type Mutant Stock; Transgenic; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Mating System +/+ sibling x Hemizygote (Female x Male) 18-APR-08 Species laboratory mouse Generation ?+F6 (18-NOV-09)
Generation DefinitionsDonating Investigator Nicholas Gaiano, Johns Hopkins Univ School of Medicine Description
Mice homozygous for the Notch reporter transgene are viable, fertile, normal in size, and do not display any behavioral abnormalities. Enhanced green fluorescent protein (EGFP) expression is present in a wide variety of hemizygous cell/tissue types during development and in the adult, including enriched hematopoietic stem cell (HSC) populations. The location of EGFP expression is consistent with Notch signaling pathway elements/genes and appears to faithfully reflect canonical (CBF1-mediated) Notch activity. With respect to hematopoiesis, low expression is shown in fully differentiated cells of the peripheral lymphoid organs (blood and spleen). Isolated HSC retain their ability to differentiate. Mice expressing this Notch reporter transgene may be useful in studying HSC populations and other cell types utilizing the Notch, CBF1, or Wnt signaling pathways. As immature (double negative [DN]) thymocytes have differential expression patterns as they progress from DN1-DN4, these mice may also be useful in thymocyte maturation studies.In addition to the above description, the donating investigator confirms the following characteristics: EGFP expression can be upregulated in transgenic mouse embryonic fibroblasts after infection with activated Notch1-3, or an activated form of CBF1 (CBF1-VP16). In addition, EGFP expression can be inhibited by shRNA-mediated knockdown of CBF1, or by treatment with a gamma-secretase inhibitor. In the developing nervous system, EGFP is expressed in the germinal zone of the brain and spinal cord, and can be used to enrich for neural progenitors by fluorescence-activated cell sorting (FACS). EGFP expression is also present in the adult nervous system, in germinal zones, and also in mature neurons. Isolated hematopoietic stem cells (HSC) retain their ability to differentiate. This line has also been used in published studies examining angiogenesis and hair follicle formation.
Development
An enhanced green fluorescent protein (EGFP) sequence was placed under the control of 4 tandem copies of the C promoter (Cp) binding factor 1 (CBF1) binding site consensus sequence (derived from Epstein Barr Virus) upstream of the SV40 basal promoter. This construct was injected into (B6SJLF1/J x B6SJLF1/J) pronuclei. Male founder #25 was obtained and bred with a female CD1 mouse. Hemizygous mice were bred with CD1 for three generations before being made homozygous. Homozygotes were bred an additional two generations before arriving at The Jackson Laboratory. Upon arrival, transgenic mice were bred to C57BL/6J inbred mice for at least one generation to establish the colony.
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Fluorescent Protein Strains
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Strains carrying other alleles of GFP
View Strains carrying other alleles of GFP (181 strains)
Fluorescent Proteins/lacZ Systems
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Research Applications
This mouse can be used to support research in many areas including:
GFP relatedCell Biology Research
Signal Transduction
Developmental Biology Research
Internal/Organ Defects
hematopoietic defects
Lymphoid Tissue Defects
hematopoietic defects
Hematological Research
Hematopoietic Defects
Immunology and Inflammation Research
Lymphoid Tissue Defects
hematopoietic development
Neurobiology Research
Fluorescent protein expression in neural tissue
Research Tools
Developmental Biology Research
transplantation marker for embryonic and adult tissue
Fluorescent Proteins
Genetics Research
Tissue/Cell Markers
Tissue/Cell Markers: cell marker for bone marrow transplantation
Tissue/Cell Markers: multiple
Immunology and Inflammation Research
Neurobiology Research
cell marker
Research Tools
Fluorescent Proteins
| Allele Symbol | Tg(Cp-EGFP)25Gaia | ||
|---|---|---|---|
| Allele Name | transgene insertion 25, Nicholas R Gaiano | ||
| Allele Type | Transgenic (Reporter) | ||
| Common Name(s) | TNR; transgenic Notch reporter; | ||
| Mutation Made By | Nicholas Gaiano, Johns Hopkins Univ School of Medicine | ||
| Strain of Origin | (C57BL/6J x SJL/J)F2 | ||
| Site of Expression | Widespread expression during development and in the adult, including enriched hematopoietic stem cell (HSC) populations. As immature (double negative [DN]) thymocytes have differential expression patterns as they progress from DN1-DN4, these mice may also be useful in thymocyte maturation studies. | ||
| 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 | C, C promoter (Cp) binding factor 1 (CBF1) binding site consensus sequence (derived from Epstein Barr Virus), | ||
| General Note | EGFP is expressed from this transgene when the promoter is activated by binding to the upstream CBF1 binding elements of a complex comprising CBF1, the transcriptional coactivator mastermind like 1 (Drosophila)(MML1), and the intracellular cleavage product resulting from interaction of the transmembrane Notch receptor with ligand. | ||
| Molecular Note | The transgene comprises four tandem copies of the C-promoter binding factor 1 (CBF1/RBPSUH) binding site of the Epstein-Barr virus major latency C promoter (Cp), followed by the SV40 minimal promoter and the coding sequence for enhanced green fluorescentprotein. Cells of mice bearing this transgene express green fluorescent protein when the Notch signaling pathway is activated. [MGI Ref ID J:96367] | ||
| Gene Symbol and Name | Tg(Cp-EGFP)25Gaia, transgene insertion 25, Nicholas R Gaiano | ||
| Chromosome | UN | ||
| Gene Common Name(s) | TNR; transgenic Notch reporter; | ||
Genotyping Protocols
Fluorescent Proteins (Generic GFP), Melt Curve Analysis
Fluorescent Proteins (Generic GFP), Standard PCR
Helpful Links
Genotyping resources and troubleshooting
Duncan AW; Rattis FM; DiMascio LN; Congdon KL; Pazianos G; Zhao C; Yoon K; Cook JM; Willert K; Gaiano N; Reya T. 2005. Integration of Notch and Wnt signaling in hematopoietic stem cell maintenance. Nat Immunol 6(3):314-22. [PubMed: 15665828] [MGI Ref ID J:96367]
Tg(Cp-EGFP)25Gaia relatedButler JM; Nolan DJ; Vertes EL; Varnum-Finney B; Kobayashi H; Hooper AT; Seandel M; Shido K; White IA; Kobayashi M; Witte L; May C; Shawber C; Kimura Y; Kitajewski J; Rosenwaks Z; Bernstein ID; Rafii S. 2010. Endothelial cells are essential for the self-renewal and repopulation of Notch-dependent hematopoietic stem cells. Cell Stem Cell 6(3):251-64. [PubMed: 20207228] [MGI Ref ID J:158983]
Estrach S; Ambler CA; Lo Celso CL; Hozumi K; Watt FM. 2006. Jagged 1 is a {beta}-catenin target gene required for ectopic hair follicle formation in adult epidermis. Development 133(22):4427-38. [PubMed: 17035290] [MGI Ref ID J:115280]
Hellstrom M; Phng LK; Hofmann JJ; Wallgard E; Coultas L; Lindblom P; Alva J; Nilsson AK; Karlsson L; Gaiano N; Yoon K; Rossant J; Iruela-Arispe ML; Kalen M; Gerhardt H; Betsholtz C. 2007. Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis. Nature 445(7129):776-80. [PubMed: 17259973] [MGI Ref ID J:118598]
Jeong HW; Jeon US; Koo BK; Kim WY; Im SK; Shin J; Cho Y; Kim J; Kong YY. 2009. Inactivation of Notch signaling in the renal collecting duct causes nephrogenic diabetes insipidus in mice. J Clin Invest 119(11):3290-300. [PubMed: 19855135] [MGI Ref ID J:154589]
Kim YW; Koo BK; Jeong HW; Yoon MJ; Song R; Shin J; Jeong DC; Kim SH; Kong YY. 2008. Defective Notch activation in microenvironment leads to myeloproliferative disease. Blood 112(12):4628-38. [PubMed: 18818392] [MGI Ref ID J:143346]
Lai AY; Kondo M. 2007. Identification of a bone marrow precursor of the earliest thymocytes in adult mouse. Proc Natl Acad Sci U S A 104(15):6311-6. [PubMed: 17404232] [MGI Ref ID J:131942]
Mercher T; Raffel GD; Moore SA; Cornejo MG; Baudry-Bluteau D; Cagnard N; Jesneck JL; Pikman Y; Cullen D; Williams IR; Akashi K; Shigematsu H; Bourquin JP; Giovannini M; Vainchenker W; Levine RL; Lee BH; Bernard OA; Gilliland DG. 2009. The OTT-MAL fusion oncogene activates RBPJ-mediated transcription and induces acute megakaryoblastic leukemia in a knockin mouse model. J Clin Invest 119(4):852-64. [PubMed: 19287095] [MGI Ref ID J:149622]
Mizutani K; Yoon K; Dang L; Tokunaga A; Gaiano N. 2007. Differential Notch signalling distinguishes neural stem cells from intermediate progenitors. Nature 449(7160):351-5. [PubMed: 17721509] [MGI Ref ID J:126684]
Wu M; Kwon HY; Rattis F; Blum J; Zhao C; Ashkenazi R; Jackson TL; Gaiano N; Oliver T; Reya T. 2007. Imaging Hematopoietic Precursor Division in Real Time. Cell Stem Cell 1(5):541-554. [PubMed: 18345353] [MGI Ref ID J:131941]
Yan Q; Yao D; Wei LL; Huang Y; Myers J; Zhang L; Xin W; Shim J; Man Y; Petryniak B; Gerson S; Lowe JB; Zhou L. 2010. O-fucose modulates notch-controlled blood lineage commitment. Am J Pathol 176(6):2921-34. [PubMed: 20363915] [MGI Ref ID J:161168]
Yoon MJ; Koo BK; Song R; Jeong HW; Shin J; Kim YW; Kong YY; Suh PG. 2008. Mind bomb-1 is essential for intraembryonic hematopoiesis in the aortic endothelium and the subaortic patches. Mol Cell Biol 28(15):4794-804. [PubMed: 18505817] [MGI Ref ID J:137915]
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, these mice are bred as hemizygotes or homozygotes. Donating investigator reports that homozygous females may have slightly reduced viability/health. Mating System +/+ sibling x Hemizygote (Female x Male) 18-APR-08 Diet Information LabDiet® 5K52/5K67
| Pricing for USA, Canada and Mexico shipping destinations |
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Price (US dollars $) Gender Genotypes Provided Individual Mouse $253.20 Female or Male Hemizygous for Tg(Cp-EGFP)25Gaia
Pairs /Price (US dollars $) Pair Genotype $309.70 Hemizygous for Tg(Cp-EGFP)25Gaia x Noncarrier $309.70 Noncarrier x Hemizygous for Tg(Cp-EGFP)25Gaia Standard Supply
Repository-Live. This collection of over 1,000 specialty strains are maintained in breeding colonies sized smaller than typical production colonies. The colonies are sized based on current customer demand. Repository-Live strains are typically delivered at four to eight weeks of age but could be up to 12 weeks of age upon receipt. The colony sizes for Repository-Live strains do not generally support filling orders with stringent age specifications or orders for large volumes of mice. However, custom orders and/or larger quantities of mice may be possible. Estimated shipping dates for all orders are provided within two business days following order placement.
Supply Notes
- Usually shipped between four and eight weeks of age.
| Pricing for International shipping destinations |
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Price (US dollars $) Gender Genotypes Provided Individual Mouse $329.20 Female or Male Hemizygous for Tg(Cp-EGFP)25Gaia
Pairs /Price (US dollars $) Pair Genotype $402.70 Hemizygous for Tg(Cp-EGFP)25Gaia x Noncarrier $402.70 Noncarrier x Hemizygous for Tg(Cp-EGFP)25Gaia Standard Supply
Repository-Live. This collection of over 1,000 specialty strains are maintained in breeding colonies sized smaller than typical production colonies. The colonies are sized based on current customer demand. Repository-Live strains are typically delivered at four to eight weeks of age but could be up to 12 weeks of age upon receipt. The colony sizes for Repository-Live strains do not generally support filling orders with stringent age specifications or orders for large volumes of mice. However, custom orders and/or larger quantities of mice may be possible. Estimated shipping dates for all orders are provided within two business days following order placement.
Supply Notes
- Usually shipped between four and eight weeks of age.
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Repository-Live. This collection of over 1,000 specialty strains are maintained in breeding colonies sized smaller than typical production colonies. The colonies are sized based on current customer demand. Repository-Live strains are typically delivered at four to eight weeks of age but could be up to 12 weeks of age upon receipt. The colony sizes for Repository-Live strains do not generally support filling orders with stringent age specifications or orders for large volumes of mice. However, custom orders and/or larger quantities of mice may be possible. Estimated shipping dates for all orders are provided within two business days following order placement.
| Control | ||
|---|---|---|
| Noncarrier | ||
| 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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| phone: | 207-288-6470 |
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