Intracellular Delivery of mRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation

• 作者: Bart Vandekerckhove Glenn Goetgeluk Jelter Van Hoeck Juan C. Fraire Kevin Braeckmans Koen Raemdonck Laurens Raes Lien Van Hoecke Ranhua Xiong Rein Verbeke Stefaan De Smedt Stephan Stremersch Stijn De Munter Toon Brans Xavier Saelens
• 来源期刊: 纳微快报（英文）
• 年，卷（期）: 2020,（12）
• 页码范围: 404-420
• 页数: 17页
• 语种: 英文

摘要

Efficient and safe cell engineering by transfection of nucleic acidsremains one of the long-standing hurdles for fundamental biomedical research and many new therapeutic applications, such as CAR T cell-based therapies. mRNA has recently gained increasing attention as a more safe and versatile alternative tool over viral- or DNA transposon-based approaches for the generation of adop-tive T cells. However, limitations associated with existing nonviral mRNA deliv-ery approaches hamper progress on genetic engineering of these hard-to-transfect immune cells. In this study, we demonstrate that gold nanoparticle-mediated vapor nanobubble (VNB) photoporation is a promising upcoming physical transfection method capable of delivering mRNA in both adherent and suspension cells. Initial transfection experiments on HeLa cells showed the importance of transfection buffer and cargo concentration, while the technology was furthermore shown to be effective for mRNA delivery in Jurkat T cells with transfection efficiencies up to 45％. Importantly, compared to electroporation, which is thereference technology for nonviral transfection of T cells, a fivefold increase in the number of transfected viable Jurkat T cells was observed. Altogether, our results point toward the use of VNB photoporation as a more gentle and efficient technology for intracellular mRNA delivery in adherent and suspension cells, with promising potential for the future engineering of cells in therapeutic and fundamental research applications.