Recombinase-activating gene 1-associated expression of the myelin basic protein 1-11-specific transgenic T-cell...

We pursued a breeding strategy intended to generate disease-resistant mice with exclusive expression of the H-2(u)-restricted myelin basic protein (MBP) 1-11 peptide-specific transgenic (Tg) T-cell receptor (TCR) on the T-cell-deficient RAG1KO (H-2(b)) background. Utilizing specific screening assays for the offspring, analyses of the F1 intercross and subsequent crosses revealed that the TgTCR-associated clonotypic marker detected by the 3H12 mAb could be found only in association with the H-2(b) homozygous background in offspring possessing a functional rag1 gene. Moreover, expression of the MBP-specific TgTCR could not be found in H-2(b) homozygous offspring that were RAG1 deficient (rag1(-/-)). PCR analysis of genomic DNA from these 3H12-negative offspring verified the presence of the TCR transgenes. Thus, the presence of a functional rag1 gene was required for the expression of the MBP-specific TgTCR on the H-2(b) background. Given the role for RAG1, the results have important implications for T-cell repertoire development.