Rapid analysis of T-cell selection in vivo using T cell-receptor retrogenic mice

Nat Methods. 2006 Mar;3(3):191-7. doi: 10.1038/nmeth858.


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-linked multicistronic 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, OT-II [corrected] and AND). We also show that retrogenic mice expressing male antigen-specific TCRs (HY, MataHari and Marilyn) facilitated the analysis of positive and negative selection in female and male 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.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Female
  • Flow Cytometry
  • Gene Transfer Techniques*
  • Genetic Vectors / genetics
  • Male
  • Mice
  • Mice, Transgenic
  • Receptors, Antigen, T-Cell / drug effects
  • Receptors, Antigen, T-Cell / genetics*
  • Receptors, Antigen, T-Cell / immunology
  • Retroviridae / genetics
  • Sensitivity and Specificity
  • T-Lymphocytes / immunology*
  • Transduction, Genetic / methods*
  • Viral Proteins / pharmacology


  • Receptors, Antigen, T-Cell
  • Viral Proteins
  • virus protein 2A