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To redirect T cells toward target cells, they can be engineered ex vivo to express antigen-specific T cell receptors (TCRs), creating TCR-engineered T cells (TCR-T). In this chapter, we provide a streamlined protocol for generating autologous mRNA-electroporated, viral antigen-specific, immunosuppressive drug-resistant armored (IDRA) TCR-T cell therapy products in a laboratory setting. We also discuss the advantages of transient mRNA electroporation over permanent genetic modification technologies for our specific application. Finally, we present a protocol to evaluate the in vitro and in vivo functionality of our TCR-T cell therapy product.

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mRNA-Based Engineering of Viral Antigen-Specific TCR-T Cells

  • 实验方案

  • 发表日期：

    2025年

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  • He Shan,
  • Antonio Bertoletti,
  • Anthony Tanoto Tan

摘要

To redirect T cells toward target cells, they can be engineered ex vivo to express antigen-specific T cell receptors (TCRs), creating TCR-engineered T cells (TCR-T). In this chapter, we provide a streamlined protocol for generating autologous mRNA-electroporated, viral antigen-specific, immunosuppressive drug-resistant armored (IDRA) TCR-T cell therapy products in a laboratory setting. We also discuss the advantages of transient mRNA electroporation over permanent genetic modification technologies for our specific application. Finally, we present a protocol to evaluate the in vitro and in vivo functionality of our TCR-T cell therapy product.