B6;129S-Gt(ROSA)26Sor^{tm1(NOTCH3*R1031C)Sat}/Mmjax

Stock number: 013206
Research areas: Cardiovascular Research, Research Tools

Description

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.

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.

Allele

Symbol: Gt(ROSA)26Sor^{tm3(NOTCH3*R1031C)Sat}
Name: targeted mutation 3, Spyros Artavanis-Tsakonas
MGI accession ID: MGI:5007799
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence; Humanized sequence
Synonyms: Gt(ROSA)26Sor^{tm1(NOTCH3*R1031C)Sat}; NOTCH3^R1031C
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: 129S6/SvEvTac
Expressed genes: NOTCH3,notch 3,human; GFP,Green Fluorescent Protein,
Site of expression: EGFP and human NOTCH3 will be expressed in cells where promoters driving Cre recombinase are expressed.
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.