Modification of specificity of T cells for the use in adoptive transfer (CAR- or TCR-redirected T cells) has revolutionized the therapy of liquid tumors and some infectious diseases. However, several obstacles are still hampering the efficacy of such potent therapy, hence concurrent modification of the function is also required to obtain successful results. Here we show the use of splice-switching antisense oligonucleotides (SSOs) as a tool to transiently modify T cell function. We demonstrate the possibility to transfect SSOs and an exogenous TCR into primary human T cells in the same electroporation reaction, without affecting viability and function of the transfected T lymphocytes. Moreover, we show that SSOs targeting T cell-specific mRNAs induce the skipping of the targeted exons, and the reduction of the protein and consequent modification of T cell function. This technical work paves the way to the use of SSOs in immune cells, not only for the knockdown of the functional isoform of the targeted proteins, but also for the protein manipulation by elimination of specific domains encoded by targeted exons.

中文翻译：

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剪接转换反义寡核苷酸作为产生装甲重定向 T 细胞的靶向内在工程工具

用于过继转移（CAR 或 TCR 重定向的 T 细胞）的 T 细胞特异性的修饰已经彻底改变了液体肿瘤和一些传染病的治疗。然而，一些障碍仍然阻碍了这种有效疗法的功效，因此还需要同时修改功能以获得成功的结果。在这里，我们展示了使用剪接转换反义寡核苷酸 (SSO) 作为瞬时修改 T 细胞功能的工具。我们证明了在相同的电穿孔反应中将 SSO 和外源性 TCR 转染到原代人 T 细胞中的可能性，而不影响转染的 T 淋巴细胞的活力和功能。此外，我们表明靶向 T 细胞特异性 mRNA 的 SSO 诱导靶向外显子的跳跃，以及蛋白质的减少和随之而来的 T 细胞功能的改变。这项技术工作为在免疫细胞中使用 SSO 铺平了道路，不仅用于击倒目标蛋白质的功能异构体，而且还用于通过消除由目标外显子编码的特定域来进行蛋白质操作。