STOCK Tg(tetO-MAML1*/EGFP)2Akar/J

Stock No:: 017918 |; TetOS-dnMAML-GFP

Also Known As: TetOS-dnMAML-GFP

Tet^OS-dnMAML-GFP transgenic mice allow Tet-Off/Tet-On expression of a dominant negative form of MAML, and may be may be useful for temporally-controlled, tissue-specific blockade of Notch signaling.

Donating Investigator

Aly Karsan, University of British Columbia, British Columbia Cancer Research Centre

Genetic overview

Genetic Background	Generation

Tg(tetO-MAML1*/EGFP)2Akar

Allele Type
Transgenic (Dominant negative, Inducible, Inserted expressed sequence, Reporter)

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Research Applications

Research Tools
Neurobiology Research
Developmental Biology Research
Cell Biology Research

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Base Price

Starting at:

$2,595.00 Domestic price Cryo Recovery

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Details

Detailed Description

The Tet^OS-dnMAML-GFP transgene has the Tet response element (TRE) upstream of a cDNA sequence encoding a dominant-negative mutant variant of human Mastermind-like 1 (dnMAML) fused in-frame at its C-terminus to an enhanced green fluorescent protein (EGFP). The dnMAML-GFP fusion protein [also called DNMAML1-GFP or MAML1(13-74)-GFP] interrupts transactivation/nuclear activity of all four Notch receptors by binding the Notch-CSL [CBF1/Su(H)/Lag2 protein] transcriptional complex, but does not bind the coactivators required for transcription of target genes.

Mice homozygous for the Tet^OS-dnMAML-GFP transgene are viable and fertile with no reported phenotypic abnormalities. When bred with other mice expressing tetracycline-controlled transactivator protein (tTA) or reverse tetracycline-controlled transactivator protein (rtTA), dnMAML-GFP expression in the resulting double mutant offspring can be regulated with tetracycline or its analog doxycycline (dox). When tTA or rtTA(+dox) is present, dnMAML expression blocks the transactivation/nuclear activity of all four Notch genes by binding to the Notch-CSL [CBF1/Su(H)/Lag2 protein] transcriptional complex (preventing Notch-CSL from binding its endogenous coactivator, MAML), and results in a Notch quadruple knockout in the tTA-expressing cells. GFP expression be observed by direct/native fluorescence and antibody staining/immunohistochemistry (the donating investigator did not attempt mRNA (in situ) hybridization). As designed, Tet^OS-dnMAML-GFP transgenic mice have no reported levels of dnMAML-GFP expression in the absence of tTA.

These Tet^OS-dnMAML-GFP transgenic mice allow Tet-Off/Tet-On expression of a dominant negative form of MAML, and may be may be useful for temporally-controlled, tissue-specific blockade of Notch signaling. For example, when bred to a strain expressing tTA in blood vessel endothelial cells (VE^tTA: Stock No. 013585), the resulting double mutant mice are useful for identifying Notch target genes in endothelial-to-mesenchymal transdifferentiation in cardiac development.

Development

The Tet^OS-dnMAML-GFP transgene was designed with a TetOS promoter followed by a cDNA sequence encoding the dnMAML-GFP fusion protein and an SV40 polyA sequence. The TetOS promoter (Tet response element or TRE) contains seven copies of the 42-bp tet operator sequence (tetO) just upstream of a minimal CMV promoter. The dnMAML-GFP cDNA sequence encodes an N-terminal, 62 amino acid (amino acids 13-74), dominant-negative mutant variant of human Mastermind-like 1 (dnMAML) fused in-frame at its C-terminus to an enhanced green fluorescent protein (EGFP). The ~3.0 kbp Tet^OS-dnMAML-GFP transgene was injected into the pronucleus of ICR oocytes fertilized by ICR males. Founder mice were bred with ICR mice to establish founder line 2. The colony was maintained by breeding Tet^OS-dnMAML-GFP transgenic mice with ICR mice for several generations, and then by breeding Tet^OS-dnMAML-GFP transgenic mice with CD-1 outbred mice for several generations prior to sending males to The Jackson Laboratory Repository in 2013. Upon arrival, males were used to cryopreserve sperm. To establish the living mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J female mice (Stock No. 000664).

Expression Data

Expressed Gene	MAML1, mastermind like transcriptional coactivator 1, human
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	These transgenic mice allow Tet-Off/Tet-On expression of a dominant negative form of MAML, and may be may be useful for temporally-controlled, tissue-specific blockade of Notch signaling. For example, when bred to a strain expressing tTA in blood vessel endothelial cells the resulting double mutant mice are useful for identifying Notch target genes in endothelial-to-mesenchymal transdifferentiation in cardiac development.

Control Suggestions

Noncarrier ()

Additional Information

Considerations for Choosing Controls

Selected References

Chang L; Noseda M; Higginson M; Ly M; Patenaude A; Fuller M; Kyle AH; Minchinton AI; Puri MC; Dumont DJ; Karsan A. 2012. Differentiation of vascular smooth muscle cells from local precursors during embryonic and adult arteriogenesis requires Notch signaling. (18): 6993-8. PubMed: 22509029 MGI: 185009
Chang AC; Fu Y; Garside VC; Niessen K; Chang L; Fuller M; Setiadi A; Smrz J; Kyle A; Minchinton A; Marra M; Hoodless PA; Karsan A. 2011. Notch initiates the endothelial-to-mesenchymal transition in the atrioventricular canal through autocrine activation of soluble guanylyl cyclase. (2): 288-300. PubMed: 21839921 MGI: 185986

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Genetics

Tg(tetO-MAML1*/EGFP)2Akar

Allele Symbol: Tg(tetO-MAML1*/EGFP)2Akar

Allele Name	transgene insertion 2, Aly Karsan
Allele Type	Transgenic (Dominant negative, Inducible, Inserted expressed sequence, Reporter)
Allele Synonym(s)	Tet^OS-dnMAML-GFP; TetOS-dnMAML; dnMAML-GFP
Gene Symbol and Name	Tg(tetO-MAML1*/EGFP)2Akar, transgene insertion 2, Aly Karsan
Gene Synonym(s)
Promoter	tetO, tet operator,
Expressed Gene	MAML1, mastermind like transcriptional coactivator 1, human
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	These transgenic mice allow Tet-Off/Tet-On expression of a dominant negative form of MAML, and may be may be useful for temporally-controlled, tissue-specific blockade of Notch signaling. For example, when bred to a strain expressing tTA in blood vessel endothelial cells the resulting double mutant mice are useful for identifying Notch target genes in endothelial-to-mesenchymal transdifferentiation in cardiac development.
Strain of Origin	ICR
Chromosome	UN
Molecular Note	The Tet-dnMAML-GFP transgene was designed with a TetOS promoter followed by a cDNA sequence encoding the dnMAML-GFP fusion protein and an SV40 polyA sequence. The TetOS promoter (Tet response element or TRE) contains seven copies of the 42-bp tet operator sequence (tetO) just upstream of a minimal CMV promoter. The dnMAML-GFP cDNA sequence encodes an N-terminal, 62 amino acid (amino acids 13-74), dominant-negative mutant variant of human Mastermind-like 1 (dnMAML) fused in-frame at its C-terminus to an enhanced green fluorescent protein (EGFP).

Disease/Phenotype

Disease Terms

Research Areas By Genotype

This mouse can be used to support research in many areas including:

Genotype: GFP related

Research Tools
- Fluorescent Proteins

Research Tools
- Tet Expression Systems
  - tTA/rtTA Responsive Strains
- Genetics Research
  - Mutagenesis and Transgenesis: transcriptional activation
  - Mutagenesis and Transgenesis: Tetop Tet System
  - Tissue/Cell Markers: endothelial cells
  - Tissue/Cell Markers: multiple
- Reproductive Biology Research
  - Tetop Tet System
- Fluorescent Proteins
- Cancer Research
  - Tetop Tet System
- Neurobiology Research
  - Tetop Tet System
- Developmental Biology Research
- Cardiovascular Research
  - Tetop Tet System
Neurobiology Research
- Fluorescent protein expression in neural tissue
- Tet Expression System
  - tTA/rtTA Responsive Strains
Developmental Biology Research
- Internal/Organ Defects
Cell Biology Research
- Signal Transduction
- Transcriptional Regulation

Mammalian Phenotype Terms by Genotype

References

Chang L; Noseda M; Higginson M; Ly M; Patenaude A; Fuller M; Kyle AH; Minchinton AI; Puri MC; Dumont DJ; Karsan A. 2012. Differentiation of vascular smooth muscle cells from local precursors during embryonic and adult arteriogenesis requires Notch signaling. (18): 6993-8. PubMed: 22509029 MGI: 185009
Chang AC; Fu Y; Garside VC; Niessen K; Chang L; Fuller M; Setiadi A; Smrz J; Kyle A; Minchinton A; Marra M; Hoodless PA; Karsan A. 2011. Notch initiates the endothelial-to-mesenchymal transition in the atrioventricular canal through autocrine activation of soluble guanylyl cyclase. (2): 288-300. PubMed: 21839921 MGI: 185986

Additional - Tg(tetO-MAML1*/EGFP)2Akar

Patenaude A; Fuller M; Chang L; Wong F; Paliouras G; Shaw R; Kyle AH; Umlandt P; Baker JH; Diaz E; Tong J; Minchinton AI; Karsan A. 2014. Endothelial-specific Notch blockade inhibits vascular function and tumor growth through an eNOS-dependent mechanism. (9): 2402-11. PubMed: 24599126 MGI: 211832

Technical Support

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Genotyping Protocols
- High Resolution Melting: Tg(tetO-MAML1*/EGFP)2Akar
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, transgenic mice may be bred with wildtype (noncarrier) mice from the colony.
- Additional Breeding and Husbandry Support

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

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Also Known As: TetOS-dnMAML-GFP

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Tg(tetO-MAML1*/EGFP)2Akar

Allele Symbol: Tg(tetO-MAML1*/EGFP)2Akar

Disease Terms

Research Areas By Genotype

This mouse can be used to support research in many areas including:

Genotype: GFP related

Mammalian Phenotype Terms by Genotype

References

Additional - Tg(tetO-MAML1*/EGFP)2Akar

Genotyping Protocols

Breeding Considerations

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability