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. 2025:2965:439-454.
doi: 10.1007/978-1-0716-4742-4_22.

Developing Aptamer-Targeted mRNA for Immunotherapy

Alexandra R Paul  1   2 , Khuloud T Al-Jamal  3   4
Affiliations

Developing Aptamer-Targeted mRNA for Immunotherapy

Alexandra R Paul et al. Methods Mol Biol. 2025.

Abstract

Selective protein expression plays a crucial role in mRNA therapeutics. In this chapter, we present a novel T-cell-targeting aptamer drug delivery platform that enables efficient mRNA delivery to immune cells for immunotherapy. We outline protocols for aptamer selection, aptamer-mRNA conjugation, characterization, and transfection into T-cells. Our results show that the OX40 mRNA-conjugated T-cell aptamer enhances mRNA transfection in murine ex vivo splenocyte T-cells, achieving a fourfold increase in CD4+ cells and a sevenfold increase in CD8+ cells compared to untargeted OX40 mRNA. This platform can be customized for other targets via aptamer selection, facilitating the development of enhanced mRNA therapies.

Keywords: Aptamers; Flow cytometry; Gel electrophoresis; Immunotherapy; Nucleic acids; Transfection; mRNA delivery.

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