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Raes L, Stremersch S, Fraire JC, et al. Intracellular Delivery of mRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation. Nanomicro Lett. 2020;12(1):185doi: 10.1007/s40820-020-00523-0.
Raes, L., Stremersch, S., Fraire, J. C., Brans, T., Goetgeluk, G., De Munter, S., Van Hoecke, L., Verbeke, R., Van Hoeck, J., Xiong, R., Saelens, X., Vandekerckhove, B., De Smedt, S., Raemdonck, K., & Braeckmans, K. (2020). Intracellular Delivery of mRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation. Nano-micro letters, 12(1), 185. https://doi.org/10.1007/s40820-020-00523-0
Raes, Laurens, et al. "Intracellular Delivery of mRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation." Nano-micro letters vol. 12,1 (2020): 185. doi: https://doi.org/10.1007/s40820-020-00523-0
Raes L, Stremersch S, Fraire JC, Brans T, Goetgeluk G, De Munter S, Van Hoecke L, Verbeke R, Van Hoeck J, Xiong R, Saelens X, Vandekerckhove B, De Smedt S, Raemdonck K, Braeckmans K. Intracellular Delivery of mRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation. Nanomicro Lett. 2020 Sep 27;12(1):185. doi: 10.1007/s40820-020-00523-0. PMID: 34138203; PMCID: PMC7770675.
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Nanomicro Lett. 2020 Sep 27;12(1):185. doi: 10.1007/s40820-020-00523-0.

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

Nano-micro letters

Laurens Raes, Stephan Stremersch, Juan C Fraire, 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

Affiliations

  1. Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000, Ghent, Belgium.
  2. Cancer Research Institute Ghent (CRIG), 9000, Ghent, Belgium.
  3. Department of Diagnostic Sciences, Ghent University, 9000, Ghent, Belgium.
  4. VIB-UGent Center for Medical Biotechnology, 9052, Ghent, Belgium.
  5. Department of Biomedical Molecular Biology, Ghent University, 9000, Ghent, Belgium.
  6. Department of Biochemistry and Microbiology, Ghent University, 9000, Ghent, Belgium.
  7. Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000, Ghent, Belgium. [email protected].
  8. Cancer Research Institute Ghent (CRIG), 9000, Ghent, Belgium. [email protected].

PMID: 34138203 PMCID: PMC7770675 DOI: 10.1007/s40820-020-00523-0

Abstract

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 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 adoptive T cells. However, limitations associated with existing nonviral mRNA delivery 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 the reference 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.

Keywords: Gold nanoparticles; Optoporation; Photoporation; Transfection; Vapor nanobubbles; mRNA

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