Light triggered nanoscale biolistics for efficient intracellular delivery of functional macromolecules in mammalian cells

Abstract

Biolistic intracellular delivery of functional macromolecules makes use of dense microparticles which are ballistically fired onto cells with a pressurized gun. While it has been used to transfect plant cells, its application to mammalian cells has met with limited success mainly due to high toxicity. Here we present a more refined nanotechnological approach to biolistic delivery with light-triggered self-assembled nanobombs (NBs) that consist of a photothermal core particle surrounded by smaller nanoprojectiles. Upon irradiation with pulsed laser light, fast heating of the core particle results in vapor bubble formation, which propels the nanoprojectiles through the cell membrane of nearby cells. We show successful transfection of both adherent and non-adherent cells with mRNA and pDNA, outperforming electroporation as the most used physical transfection technology by a factor of 5.5–7.6 in transfection yield. With a throughput of 104-105 cells per second, biolistic delivery with NBs offers scalable and highly efficient transfections of mammalian cells.

Photothermal Nanomaterial-Mediated Photoporation

“…C.ShaabaniE.SharifiaghdamM.RombautM.HinnekensC.HuaD.RamonJ.RaesL.Bolea-FernandezE.BransT.VanhaeckeF.BorghgraefP.HuangC.SauvageF.VanhaeckeT.De KockJ.XiongR.De SmedtS.BraeckmansK. Fraire, J. C. Shaabani, E. Sharifiaghdam, M. Rombaut, M. Hinnekens, C. Hua, D. Ramon, J. Raes, L. Bolea-Fernandez, E. Brans, T. Vanhaecke, F. Borghgraef, P. Huang, C. Sauvage, F. Vanhaecke, T. De Kock, J. Xiong, R. De Smedt, S. Braeckmans, K. 10.1038/s41467-022-29713-72022131996 …”

“…The intracellular delivery of macromolecules becomes increasingly difficult for increasing size due to slower diffusional mobility and steric hindrance at the pores. To increase the delivery efficiency of large cargo molecules such as mRNA and pDNA, we developed a composite nanostructure, called nanobombs, to create larger pores in the cell membrane . The light-triggered nanobombs were made of a photothermal core particle coated with smaller nanoparticles which function as nanoprojectiles (Figure a).…”

“…(b) Schematic overview of intracellular delivery by membrane permeabilization with photothermal nanofibers. Reproduced with permission from refs and . Copyrights 2021 and 2022 Nature Publishing Group.…”

“…To increase the delivery efficiency of large cargo molecules such as mRNA and pDNA, we developed a composite nanostructure, called nanobombs, to create larger pores in the cell membrane. 4 The light-triggered nanobombs were made of a photothermal core particle coated with smaller nanoparticles which function as nanoprojectiles (Figure 6a).…”

Quantized energy harvesting in vibrating maglev graphite driven by terahertz waves

“…Electromagnetic wave-driven micro-nano motors have important applications in micro-nano-mechanical systems and energytransfer systems because they have the advantages of remote control, no waste, and fast conversion. [1][2][3] At present, most research on light-driven micro-nano motors focuses on the driving source of optical pressure or optical momentum, [4][5][6][7][8][9][10][11][12][13] photothermal materials, [14][15][16][17] photothermal-magnetic materials, 18,19 hyperbolic metamaterials, 20 light-driven actuating materials, 21 bioinspired gradient design, 22 photocatalytic driving, [23][24][25][26][27][28][29] etc. However, the motion of micro-nano motors is susceptible to external parameters.…”