Tcra^Ln2Sdou

This allele is generated using a somatic cell nuclear transfer (SCNT). Male agouti (C57BL/6 x 129)F1 mice were immunized with palmitoylated TRYP1 (tyrosinase-related protein or TRP1) peptide, anti-CD40 antibody and poly-IC to create a population of TRP1 specific lymphocytes. This peptide-based vaccine elicits a robust population of tetramer+ CD8+ T cells that can reject established B16 tumors. Six to eight days post-immunization, TRP1 tetramer+ populations were observed in the spleen. Using flow cytometry, CD8+ TRP1 tetramer+ populations were isolated and individual donor nuclei selected and placed into an enucleated oocyte. Cells were developed to the blastocyst stage and used to generate embryonic stem (ES) cells. ES cells were injected into recipient blastocysts, and blastocysts implanted into pseudopregnant females. Tcra was sequenced and identified as V6-5*04 or V6D-5*01 or V6N-5*01 and J26*01 and propagated. (J:254784)