B6.Cg-Tg(CMV-CASP3)17Edge/J

Stock number: 016908
Research areas: Apoptosis Research, Cell Biology Research, Diabetes and Obesity Research, Neurobiology Research, Research Tools

Description

Mos-iCsp3 transgenic mice have a CMV promoter directing expression of a floxed-lacZ-STOP cassette followed by two tandem FK506-binding sites (Fvs) and a downstream human Caspase 3 (Casp3) gene. These mice may be useful for cell ablation of specific cell types and as a model of degenerative diseases such as Parkinson's Disease, type 1 diabetes, alopecia, and deafness.

Detailed description

Mos-iCsp3 transgenic mice have a CMV promoter directing expression of a floxed-lacZ-STOP cassette followed by two tandem FK506-binding sites (Fvs) and a downstream human Caspase 3 (Casp3) gene. Hemizygous Mos-iCsp3 mice are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Caspase 3 is a member of the cysteine-aspartic acid protease family of enzymes which are integral to apoptosis pathways. Inactive until cleaved by an initiator enzyme, Caspase 3 is processed at conserved aspartic residues and is activated by the formation of dimers. In this transgenic strain, a STOP cassette is present, flanked by a loxH site and a loxP site, preventing inducible Caspase 3 expression and allowing for widespread lacZ staining. LacZ staining in founder line 17 is seen in the spinal cord, muscle, pancreas, skin, and portions of the brain and skull. The presence of both a loxH site and a loxP site causes reduced cre efficiency, resulting in mosaic expression of Caspase 3 after cre-mediated recombination. This inducible Caspase 3 is bound to 2 tandem FK506-binding sites which act to bring 2 monomers together after induction by the FK506 analogue, AP20187. This dimerization activates Caspase 3 in cre-expressing cells, resulting in cell death. These mice may be useful for cell ablation of specific cell types and as a model of degenerative diseases such as Parkinson's Disease, type 1 diabetes, alopecia, and deafness.

Development

The Mos-iCsp3 transgene was designed with (from 5' to 3') a CMV promoter, a loxH site, a β-galactosidase (lacZ) gene, a V5-tag (derived from the paramyxovirus of simian virus 5 (SV5)), a 6x-histidine-tag, a polyadenylation (polyA) signal, and a loxP site. This floxed-lacZ-STOP cassette was followed by a myristoylation sequence, two tandem FK506-binding sites (Fvs), a human Caspase 3 (Casp3) gene, a haemagglutinin (HA) tag, and a second (polyA) signal. The transgene was microinjected into fertilized C57BL/6 X C3H/HeN F1 oocytes, and mice from founder line 17 were bred to C57BL/6J mice for at least 8 generations to establish a colony. Upon arrival at The Jackson Laboratory, transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664).

Allele

Symbol: Tg(CMV-CASP3)17Edge
Name: transgene insertion 17, Albert Edge
MGI accession ID: MGI:5014208
Generation method: Transgenic
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(CMV-CASP3)17Edge
Marker name: transgene insertion 17, Albert Edge
Chromosome: UN
Strain of origin: (C57BL/6 x C3H/HeN)F1
Expressed genes: CASP3,caspase 3,human
Molecular note: The Mos-iCsp3 transgene was designed with (from 5' to 3') a CMV promoter, a loxH site, a beta-galactosidase (lacZ) gene, a V5-tag (derived from the paramyxovirus of simian virus 5 (SV5)), a 6x-histidine-tag, a polyadenylation (polyA) signal, and a loxP . This floxed-lacZ-STOP cassette was followed by a myristoylation sequence, two tandem FK506-binding sites (Fvs), a human Caspase 3 (CASP3) gene, a haemagglutinin (HA) tag, and a second (polyA) signal. Lines 14 and 17 were generated.