Overview

In this ROSA NOTCH3-1031 knockin strain, the human NOTCH3 gene is altered by the CADASIL-associated R1031C mutation and expression is blocked by an upstream loxP-flanked STOP sequence. Cre-mediated excision results in expression of the mutated NOTCH3 and GFP targeted to the nucleus. Expression in vascular smooth muscle cells is characterized by cell degeneration and the presence of granular osmiophilic deposits. These mice allow inducible expression of a human mutation associated with ischemic stroke.

Donating Investigator

Spyros Artavanis-Tsakonas, Harvard Medical School

Genetic overview

Genetic Background	Generation

Gt(ROSA)26Sor^{tm3(NOTCH3R1031C)Sat}*

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence, Humanized sequence)	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

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Research Applications

Research Tools
Cardiovascular Research

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Details

Detailed Description

Homozygotes: Homozygous ROSA NOTCH3-1031 mice are viable, fertile, with the CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy)-associated R1031C mutation inserted into the Gt(ROSA)26Sor locus. Expression of the mutated human NOTCH3 is blocked by an upstream loxP-flanked STOP sequence. When bred to cre expressing mice, the STOP sequence is deleted in the tissues of offspring where Cre recombinase is present, resulting in mutated human NOTCH3 expression and the parallel expression of enhanced green fluorescent protein (GFP) targeted to the nucleus. Expression in vascular smooth muscle cells is characterized by cell degeneration and the presence of granular osmiophilic deposits. These mice allow inducible expression of a human mutation associated with ischemic stroke.

Heterozygote: Not evaluated

Development

A targeting vector was designed with a splice acceptor site, a loxP-flanked PGK neomycin cassette, a human NOTCH3 cDNA encoding a CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy)-associated mutation and an IRES-nuclear EGFP sequence was inserted into the GT(ROSA)26Sor locus. The CADASIL mutation is an amino acid substitution from arginine to cysteine at amino acid 1031 (R1031C). The construct was introduced into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting "1031" chimeric animals were crossed to 129S2/SvEv mice. The mice were crossed to 129S2/SvEv for an unknown number of generations and at some point to C57BL/6 mice. Upon arrival, mice were bred to 129S1/SvImJ for at least 1 generation to establish the colony.

Expression Data

Expressed Gene	NOTCH3, notch 3, human
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	EGFP and human NOTCH3 will be expressed in cells where promoters driving Cre recombinase are expressed.

Control Suggestions

Approximate Controls

Wild-type from the colony

Additional Information

Considerations for Choosing Controls

Selected References

Arboleda-Velasquez JF; Manent J; Lee JH; Tikka S; Ospina C; Vanderburg CR; Frosch MP; Rodriguez-Falcon M; Villen J; Gygi S; Lopera F; Kalimo H; Moskowitz MA; Ayata C; Louvi A; Artavanis-Tsakonas S. 2011. PNAS Plus: Hypomorphic Notch 3 alleles link Notch signaling to ischemic cerebral small-vessel disease. Proc Natl Acad Sci U S A 108(21):E128-35PubMed: 21555590 MGI: J:171887

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Genetics

Gt(ROSA)26Sor^{tm3(NOTCH3R1031C)Sat}*

Allele Symbol: Gt(ROSA)26Sor^{tm3(NOTCH3R1031C)Sat}*

Allele Name	targeted mutation 3, Spyros Artavanis-Tsakonas
Allele Type	Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)	Gt(ROSA)26Sor^{tm1(NOTCH3*R1031C)Sat}; NOTCH3^R1031C
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)
Expressed Gene	NOTCH3, notch 3, human
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	EGFP and human NOTCH3 will be expressed in cells where promoters driving Cre recombinase are expressed.
Strain of Origin	129S6/SvEvTac
Chromosome	6
Molecular Note	A targeting vector containing a splice acceptor site, a floxed PGK neomycin cassette, a human NOTCH3 cDNA with an R1031C mutation and an IRES-nuclear GFP sequence was inserted via homologous recombination.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Fluorescent Proteins
- Cardiovascular Research
  - Cre-lox System
Cardiovascular Research
- Ischemia studies

Mammalian Phenotype Terms by Genotype

References

Arboleda-Velasquez JF; Manent J; Lee JH; Tikka S; Ospina C; Vanderburg CR; Frosch MP; Rodriguez-Falcon M; Villen J; Gygi S; Lopera F; Kalimo H; Moskowitz MA; Ayata C; Louvi A; Artavanis-Tsakonas S. 2011. PNAS Plus: Hypomorphic Notch 3 alleles link Notch signaling to ischemic cerebral small-vessel disease. Proc Natl Acad Sci U S A 108(21):E128-35PubMed: 21555590 MGI: J:171887

Additional - Gt(ROSA)26Sor^{tm3(NOTCH3R1031C)Sat}* related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Generic Rosa26Sor(GFP)
- Genotyping resources and troubleshooting
Breeding Considerations

While maintaining a live colony, these mice are bred as homozygotes.
- Additional Breeding and Husbandry Support
Citation
When using the B6;129S-Gt(ROSA)26Sor^{tm1(NOTCH3*R1031C)Sat}/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #32999 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

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

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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

Terms Of Use

General Terms and Conditions

See MMRRC for Additional Conditions of Distribution

Questions about Terms of Use

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Gt(ROSA)26Sortm3(NOTCH3*R1031C)Sat

Allele Symbol: Gt(ROSA)26Sortm3(NOTCH3*R1031C)Sat

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

References

Additional - Gt(ROSA)26Sortm3(NOTCH3*R1031C)Sat related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

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

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

Gt(ROSA)26Sor^{tm3(NOTCH3R1031C)Sat}*

Allele Symbol: Gt(ROSA)26Sor^{tm3(NOTCH3R1031C)Sat}*

Additional - Gt(ROSA)26Sor^{tm3(NOTCH3R1031C)Sat}* related