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

Methods Mol Biol. 2025:2965:275-284. doi: 10.1007/978-1-0716-4742-4_13.

Abstract

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.

Keywords: Adoptive T cell transfer; IDRA; TCR-T cell therapy; T cell in vitro expansion; Virus-specific T cells; mRNA electroporation.

MeSH terms

  • Antigens, Viral* / genetics
  • Antigens, Viral* / immunology
  • Electroporation / methods
  • Genetic Engineering* / methods
  • Humans
  • Immunotherapy, Adoptive / methods
  • RNA, Messenger* / genetics
  • Receptors, Antigen, T-Cell* / genetics
  • Receptors, Antigen, T-Cell* / immunology
  • Receptors, Antigen, T-Cell* / metabolism
  • Receptors, Chimeric Antigen* / genetics
  • Receptors, Chimeric Antigen* / immunology
  • T-Lymphocytes* / immunology
  • T-Lymphocytes* / metabolism

Substances

  • Receptors, Antigen, T-Cell
  • RNA, Messenger
  • Antigens, Viral
  • Receptors, Chimeric Antigen