STOCK Th^{tm1.1(cre/ERT2)Ddg}/J

Stock number: 025614
Product name: TH-2A-CreER
Research areas: Neurobiology Research, Research Tools

Description

These mice express tamoxifen-inducible cre recombinase (cre/ERT2) under the regulatory control of the mouse Th (tyrosine hydroxylase) gene.

Detailed description

Th (tyrosine hydroxylase) encodes an enzyme that catalyzes the rate-limiting step in this synthesis of catecholamines. It converts amino acid L-tyrosine to L-3,4-dihydroxyphenylalanine (L-DOPA), a precursor to the dopamine neurotransmitter.

TH-2A-CreER mice carry tamoxifen-inducible Cre-ERT2 under the regulatory control of the mouse Th promoter and the self-cleaving porcine Teschovirus 2A (T2A) bicistronic/multicistronic peptide. When crossed with a strain bearing a loxP-flanked sequence of interest, induced excision of the floxed region by cre is consistent with that of TH in the symphathetic ganglia, dorsal root ganglia and retina.

Development

Porcine Teschovirus 2A (T2A) cleavage peptide, and Cre-ERT2 (cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain) were introduced to the 3'UTR of the mouse Th gene. A FRT-PGK-EM7-NEOpolyA-FRT-loxP cassette was placed downstream of the 3'UTR region. The mutations were created through homologous recombination in 129S6/SvEvTac-derived embryonic stem (ES) cells. Resultant chimeric mice were crossed with C57BL/6-SJL mixed background Flp mice (see Stock No. 003800) to excise the neomycin cassette, and were subsequently backcrossed to C57BL/6J mice for 3 generations by the donating lab.

Upon arrival at The Jackson Laboratory, sperm was cryopreserved. Cryorecovery/live colony development will involve C57BL/6J (Stock No. 000664).

Allele

Symbol: Th^{tm1.1(cre/ERT2)Ddg}
Name: targeted mutation 1.1, David Ginty
MGI accession ID: MGI:5569743
Generation method: Targeted
Attributes: Recombinase-expressing; Inducible
Marker symbol: Th
Marker name: tyrosine hydroxylase
Chromosome: 7
Strain of origin: 129S6/SvEvTac
Expressed genes: cre/ERT2,Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of expression: When crossed with a strain bearing a loxP-flanked sequence of interest, induced excision of the floxed region by cre is consistent with the expression of Th in the symphathetic ganglia, dorsal root ganglia and retina.
Molecular note: A T2A cleavage peptide and the cre/ERT2 fusion protein-coding sequence were introduced to the 3'UTR of the mouse Th gene. An FRT-PGK-EM7-NEOpolyA-FRT-loxP cassette was placed downstream of the 3'UTR region. Homologous recombination was achieved in 129S6/SvEvTac-derived embryonic stem cells and resultant chimeric mice were crossed with Flp mice to excise the neomycin cassette.