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

Nano-Micro Letters 2020年第12期12卷 404-420页

作者： Laurens Raes Stephan Stremersch Juan CFraire Toon Brans glenn goetgeluk Stijn De Munter Lien Van Hoecke Rein Verbeke Jelter Van Hoeck Ranhua Xiong Xavier Saelens Bart Vandekerckhove Stefaan De Smedt Koen Raemdonck Kevin Braeckmans Laboratory of General Biochemistry&Physical Pharmacy Ghent University9000 GhentBelgium Cancer Research Institute Ghent(CRIG) 9000 GhentBelgium Department of Diagnostic Sciences Ghent University9000 GhentBelgium VIB-UGent Center for Medical Biotechnology 9052 GhentBelgium Department of Biomedical Molecular Biology Ghent University9000 GhentBelgium Department of Biochemistry and Microbiology Ghent University9000 GhentBelgium

Efficient and safe cell engineering by transfection of nucleic acids remains one of the long-standing hurdles for fundamental biomedical research and many new therapeutic applications,such as CAR T cell-based *** has recently gained increasing attention as a more safe and versatile alternative tool over viral-or DNA transposon-based approaches for the generation of adoptive T ***,limitations associated with existing nonviral mRNA delivery approaches hamper progress on genetic engineering of these hard-to-transfect immune *** 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 *** 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 the reference technology for nonviral transfection of T cells,a fivefold increase in the number of transfected viable Jurkat T cells was ***,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.

关键词： Transfection mRNA Photoporation Optoporation Gold nanoparticles Vapor nanobubbles

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