Issue

Vol. 12 (2020)

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

https://doi.org/10.1007/s40820-020-00523-0
Laurens Raes
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
http://orcid.org/0000-0002-7461-7789
Stephan Stremersch
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
http://orcid.org/0000-0002-6528-8266
Juan C. Fraire
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
http://orcid.org/0000-0002-4887-2161
Toon Brans
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
Glenn Goetgeluk
Cancer Research Institute Ghent (CRIG), 9000 Ghent, Belgium
Stijn De Munter
Cancer Research Institute Ghent (CRIG), 9000 Ghent, Belgium
http://orcid.org/0000-0003-3821-0620
Lien Van Hoecke
Cancer Research Institute Ghent (CRIG), 9000 Ghent, Belgium
http://orcid.org/0000-0001-8602-4211
Rein Verbeke
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
http://orcid.org/0000-0003-1849-5411
Jelter Van Hoeck
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
Ranhua Xiong
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000, Ghent, Belgium
http://orcid.org/0000-0001-9917-8949
Xavier Saelens
VIB-UGent Center for Medical Biotechnology, 9052 Ghent, Belgium
http://orcid.org/0000-0002-3861-6965
Bart Vandekerckhove
Cancer Research Institute Ghent (CRIG), 9000 Ghent, Belgium
http://orcid.org/0000-0003-3828-4195
Stefaan De Smedt
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
Koen Raemdonck
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
Kevin Braeckmans
Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium
http://orcid.org/0000-0002-7993-6295

Corresponding Author: Kevin Braeckmans

Kevin.Braeckmans@UGent.be

Nano-Micro Letters, Vol. 12 (2020), Article Number: 185

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.


Highlights:


1 Vapor nanobubble (VNB) photoporation represents a promising physical technique for mRNA transfection of adherent and suspension cells.
2 A multitude of parameters related to the VNB photoporation procedure were optimized to enable efficient mRNA transfection.
3 VNB photoporation was found to yield five times more living, transfected Jurkat T cells as compared to electroporation, i.e., currently the standard nonviral transfection technique for T cells.

Keywords

Transfection mRNA Photoporation Optoporation Gold nanoparticles Vapor nanobubbles
Raes, Laurens, 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, and Kevin Braeckmans. 2020. “Intracellular Delivery of MRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation”. Nano-Micro Letters 12 (September):185. https://doi.org/10.1007/s40820-020-00523-0.
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