B6.Cg-7630403G23Rik^{Tg(Th-cre)1Tmd} Tg(CAG-OVAL,-GFP)1Wpeng/J

Stock number: 039164
Product name: TH-OVA (TH-Cre;OVA^Tg)
Research areas: Neurobiology Research, Research Tools

Description

TH-OVA mice express Flag-tagged ovalbumin (OVA) fused to an EGFP in neuronal tissues that express tyrosine hydroxylase (TH). These mice may be useful in studies of Parkinson's disease and other neurodegenerative disorders.

Additionally, the single mutant strain OVA^Tg (absent of Cre alleles), which allows Cre-inducible expression of OVA and GFP in other tissues, is available as Stock No. 039165.

Detailed description

Ovalbumin (OVA), the major protein component of egg white, can act as surrogate tumor antigen, useful in studies of tumor immunology and T cell responses.

These TH-OVA mice express Flag-tagged ovalbumin fused to an EGFP (green fluorescent) reporter in neuronal tissues that express tyrosine hydroxylase (TH). The rat TH promoter drives expression of Cre recombinase, mediating the excision of a loxP-flanked Stop cassette in the OVA^Tg transgene to induce expression of ovalbumin in catecholaminergic cells of the midbrain. Mid-brain tissues from TH-OVA mice can be recognized by OVA-specific CD8⁺ T cells (OTI) when pulsed with dendritic cells (DCs). Immunohistochemistry is required to visualize the GFP.

The donating laboratory does not know if compound homozygotes are viable and fertile as their genotyping assays do not distinguish homozygotes and hemizygotes. They intercross female and male animals that carry both the TH-Cre allele and the Ova^Tg allele without encountering any breeding issues.

Distribution of Stock No. 039164 was supported by the Michael J. Fox Foundation for Parkinson's Research.

Additionally, the single mutant strain OVA^Tg (absent of Cre alleles), which allows Cre-inducible expression of OVA and GFP in other tissues, is available as Stock No. 039165.

Development

To develop the Ova^Tg transgenic allele, a transgenic vector carrying a CAG-loxP-pA STOP-loxP-3xFLAG-chicken ovalbumin (MF321659.1)-IRES-GFP-polyA cassette was introduced to C57BL/6 embryos. Ova^Tg transgenic mice were intercrossed with TH-Cre (Stock No. 008601) mice to produce these compound mutant TH-OVA animals.

In brief, the TH-Cre (Stock No. 008601) transgene was created when a transgenic vector expressing Cre recombinase from the rat tyrosine hydroxylase promoter was injected into B6SJLF2 embryos. The transgene was inserted in the mouse 7630403G23Rik (RIKEN cDNA 7630403G23 gene) locus on chromosome 9 (at 33,514,690-34,139,124; Build GRCm38/mm10), creating a 624,434 bp deletion in founder line 1. Additionally, this TH-Cre transgene contains (or is closely linked to) a kanamycin resistance cassette (see Stock No. 008601). Resultant mice were backcrossed to C57BL/6NCrl and C57BL/6NJ mice. SNP analysis revealed a mix of C57BL/6J and C57BL/6N-derived polymorphisms.

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

Allele

Symbol: 7630403G23Rik^{Tg(Th-cre)1Tmd}
Name: transgene insertion 1, Ted M Dawson
MGI accession ID: MGI:3584380
Generation method: Transgenic
Attributes: Recombinase-expressing
Synonyms: Tg(Th-cre)1Tmd; TH-cre
Marker symbol: 7630403G23Rik
Marker name: RIKEN cDNA 7630403G23 gene
Chromosome: 9
Strain of origin: (C57BL/6 x SJL)F2
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: dopaminergic neurons
Molecular note: Cre recombinase is expressed under the control of the rat tyrosine hydroxylase promoter. Cre recombinase activity is detected in catecholaminergic cells. This is a representative record representing TH-cre1 - TH-cre5 all of which have identical activity. Line 1 inserted into the gene at 33514690-34139124 (Build GRCm38/mm10) resulting in a 624,434 bp deletion. Founder line 1 has a copy number of greater than 20.

Allele

Symbol: Tg(CAG-OVAL,-GFP)1Wpeng
Name: transgene insertion 1, Weiyi Peng
MGI accession ID: MGI:7580840
Generation method: Transgenic
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence; Epitope tag
Synonyms: Ova^Tg
Marker symbol: Tg(CAG-OVAL,-GFP)1Wpeng
Marker name: transgene insertion 1, Weiyi Peng
Chromosome: UN
Strain of origin: C57BL/6
Molecular note: A transgenic vector carrying a CAG-loxP-pA STOP-loxP-3xFLAG-chicken ovalbumin (OVAL)-IRES-GFP-polyA cassette was introduced to C57BL/6 embryos to create the allele. The copy number and integration site are unknown.