期刊
NATURE METHODS
卷 3, 期 3, 页码 191-197出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth858
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资金
- NCI NIH HHS [CA-21765] Funding Source: Medline
- NIAID NIH HHS [AI-52199] Funding Source: Medline
Although T-cell receptor (TCR) transgenic as well as knockout and knockin mice have had a large impact on our understanding of T-cell development, signal transduction and function, the need to cross these mice delays experiments considerably. Here we provide a methodology for the rapid expression of TCRs in mice using 2A peptide-tinked muticistronic retroviral vectors to transduce stem cells of any background before adoptive transfer into RAG-1(-/-) mice. For simplicity, we refer to these as retrogenic mice. We demonstrate that these retrogenic mice are comparable to transgenic mice expressing three commonly used TCRs (OT-I, OY-II and AND). We also show that retrogenic mice expressing mate antigen-specific TCRs (HY, MataHari and Marilyn) facilitated the analysis of positive and negative selection in female and mate mice, respectively. We examined various tolerance mechanisms in epitope-coupled TCR retrogenic mice. This powerful resource could expedite the identification of proteins involved in T-cell development and function.
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