Intracellular Delivery of mRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation
- PDF / 1,829,310 Bytes
- 17 Pages / 595.276 x 790.866 pts Page_size
- 82 Downloads / 141 Views
ARTICLE
Cite as Nano-Micro Lett. (2020) 12:185 Received: 13 May 2020 Accepted: 22 August 2020 © The Author(s) 2020
https://doi.org/10.1007/s40820-020-00523-0
Intracellular Delivery of mRNA in Adherent and Suspension Cells by Vapor Nanobubble Photoporation Laurens Raes1,2 , Stephan Stremersch1,2 , Juan C. Fraire1 , Toon Brans1,2, Glenn Goetgeluk2,3, Stijn De Munter2,3 , Lien Van Hoecke2,4,5 , Rein Verbeke1,2 , Jelter Van Hoeck1,2, Ranhua Xiong1 , Xavier Saelens4,6 , Bart Vandekerckhove2,3 , Stefaan De Smedt1,2, Koen Raemdonck1,2, Kevin Braeckmans1,2 *
HIGHLIGHTS • Vapor nanobubble (VNB) photoporation represents a promising physical technique for mRNA transfection of adherent and suspension cells. • A multitude of parameters related to the VNB photoporation procedure were optimized to enable efficient mRNA transfection. • 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.
ABSTRACT Efficient and safe cell engineering by transfection of nucleic acids
remains one of the long-standing hurdles for fundamental biomedical research and
Pulsed laser irradiation
Vapor NanoBubble (VNB) photoporation
Intracellular mRNA delivery and eGFP expression
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 adopery 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
5x Jurkat Cell Yield
tive T cells. However, limitations associated with existing nonviral mRNA deliv-
method capable of delivering mRNA in both adherent and suspension cells. Initial transfection experiments on HeLa cells showed the importance of transfection
Electroporation
VNB
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 Transfection; mRNA; Photoporation; Optoporation; Gold nanoparticles; Vapor nanobubbles * 1 2 3 4 5 6
Kevin Braeckmans, [email protected] Laboratory of General Biochemistry & Physical Pharmacy, Ghent University, 9000 Ghent, Belgium Cancer Research Institute Ghent (CRIG), 9000 Ghent, Belgium Department of Diagnosti
Data Loading...
