Spatially Resolved Cell-Fate Engineering through Nanoneedle-Mediated Optoporation

Abstract

The development of advanced tissue models demands spatially resolved control over cell fate to provide deeper insights into complex biological processes, yet precise intracellular manipulation remains a critical bottleneck in cellular engineering strategies. Current approaches for gene delivery struggle in achieving the required localized resolution to control individual cell fate, while optogenetic approaches are constrained by the requirement for complex genetic engineering and limited modes of gene regulation. Here, we show that nanoneedle-mediated optoporation enables spatially resolved intracellular delivery of nucleic acids, locally directing cell fate. Through computational modeling and experimental validation, we investigated the dependence of electromagnetic field enhancement on wavelength, nanoneedle geometry, and porosity. We demonstrated the importance of surface chemistry for optoporation efficiency, achieving optimal transfection performance through thermal carbonization. This approach achieves efficient delivery for propidium iodide, 500 kDa dextran, and siRNA, alongside a 72% mRNA transfection efficiency, resulting in sustained protein expression after 24 h. Selective SNAIL1 mRNA transfection demonstrated localized cell-fate engineering, inducing a targeted epithelial to mesenchymal transition. This porous nanoneedle system presents a versatile and flexible approach for precise genetic manipulation through electromagnetic field tunability, offering new possibilities for studying complex biological processes and disease progression through spatially resolved control of cell fate.