Shan He, Bertoletti Antonio, Tan Anthony Tanoto
Signature Research Program in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore, Singapore.
Methods Mol Biol. 2025;2965:275-284. doi: 10.1007/978-1-0716-4742-4_13.
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.
为了使T细胞靶向靶细胞，可以在体外对它们进行工程改造，使其表达抗原特异性T细胞受体（TCR），从而产生TCR工程化T细胞（TCR-T）。在本章中，我们提供了一个简化的方案，用于在实验室环境中生成自体mRNA电穿孔、病毒抗原特异性、免疫抑制药物抗性强化（IDRA）TCR-T细胞治疗产品。我们还讨论了在我们的特定应用中，瞬时mRNA电穿孔相对于永久基因修饰技术的优势。最后，我们提出了一个评估我们的TCR-T细胞治疗产品体外和体内功能的方案。
