B6.Cg-Rag1^tm1Mom Tyrp1^B-w Tg(Tcra,Tcrb)9Rest/J

Stock number: 008684
Product name: RAG1^- B^W TRP-1 TCR
Research areas: Cancer Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools

Description

These mice express a MHC class II-restricted T-cell receptor recognizing the endogenous melanocyte differentiation antigen minimal TRP-1 (tyrosinase-related protein 1) epitope. This strain is a source for melanocyte reactive CD4⁺ cells from antigen-negative animals and may be useful in studies of cancer immunology and therapeutics.

Detailed description

These mice are hemizygous for the TRP-1 transgene Tg(Tcra,Tcrb)9Rest, homozygous for the targeted mutation Rag1^tm1Mom and homozygous for the white based brown radiation induced mutation of tyrosinase-related protein 1, Tyrp1^B-w. These mutant mice express a MHC class II-restricted TCR recognizing the endogenous melanocyte differentiation antigen minimal TRP-1 (tyrosinase-related protein 1) epitope corresponding to amino acids 113 to 127. The transgene is located on the Y chromosome in founder line 9. On a RAG-deficient background, these mice are also homozygous for the Tyrp1^B-w mutation and do not produce endogenous tyrosinase-related protein 1. This strain is a source for melanocyte reactive CD4⁺ cells from antigen-negative animals and may be useful in studies of cancer immunology and therapeutics.

Development

The TRP-1 TCR was generated from immunized Tyrp1^B-w mice. Clone 7A6 expressed Valpha3.2Vbeta14 (Vα3.2 and Vβ14) TCR chains. These TCR sequences were cloned into separate vectors and co-injected into microinjected into C57BL/6 fertilized eggs. Founder line 9 was subsequently established and the transgene insertion site was found on the Y chromosome.

The Tyrp1^B-w, white based brown mutation was generated in (101/RI x CJH/RI)FI exposed to ionizing radiation and involves the inversion at the 5' end of the tyrosinase-related protein 1 gene. The mice carrying the Tyrp1^B-w mutation were backcrossed to the C57BL/6N background for 8 generations.

To generate this triple mutant strain, founder line 9 animals were crossed to mice with the Rag1^tm1Mom targeted mutation on the C57BL/6 background. The mice were then crossed to mice carrying both the Rag1^tm1Mom and Tyrp1^B-w mutations. The strain is maintained hemizygous for the TRP-1 transgene Tg(Tcra,Tcrb)9Rest, homozygous for the targeted mutation Rag1^tm1Mom and homozygous for the white based brown radiation induced mutation of tyrosinase-related protein 1, Tyrp1^B-w.

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 43 markers, on Chromosome 12, was segregating. Also, 2 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating, including the rd8 recessive mutation associated with retinal degeneration on Chromosome 1 within the Crb1 locus. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Rag1^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857241
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Rag1
Marker name: recombination activating gene 1
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt⁺
Site of expression: expression is seen in bone marrow derived cell lines.
Molecular note: A 1356 bp genomic fragment of the Rag1 gene, encoding the nuclear localization signal and the zinc-finger motif, was replaced by a neomycin cassette. A mutant transcript expressed from this allele was detected by Northern blot in bone marrow derived cell lines from homozygous mice.

Allele

Symbol: Tyrp1^B-w
Name: white based brown
MGI accession ID: MGI:1855963
Generation method: Radiation induced
Marker symbol: Tyrp1
Marker name: tyrosinase-related protein 1
Chromosome: 4
Strain of origin: (101/Rl x C3H/Rl)F1
Molecular note: This allele has a gross DNA inversion at the 5' end of the gene. No detectable transcript is produced from this allele.

Allele

Symbol: Tg(Tcra,Tcrb)9Rest
Name: transgene insertion 9, Nicholas P Restifo
MGI accession ID: MGI:3826818
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(Tcra,Tcrb)9Rest
Marker name: transgene insertion 9, Nicholas P Restifo
Chromosome: Y
Strain of origin: C57BL/6
Expressed genes: Tcra,T cell receptor alpha chain,mouse, laboratory; Tcrb,T cell receptor beta chain,mouse, laboratory
Molecular note: The TRP-1 TCR was generated from immunized Tyrp1B-w mice. Clone 7A6 expressed Valpha3.2Vbeta14 TCR chains. These TCR sequences were cloned into separate vectors and co-injected into microinjected into C57BL/6 fertilized eggs. Founder line 9 was subsequently established and the transgene insertion site was found on the Y chromosome.