Nanopores and nanostraws for cell transfection

In this talk,  I will review our work on optimizing the transfection of hard-to-transfect cells, such as stem cells and clonal beta cells, using nanostraws and nanopores. Conventional transfection methods suffer from high cytotoxicity or low transfection efficiency, which negatively affect their outcome. In contrast, nanostraw- and nanopore-electroporation is a mild approach that can achieve high transfection efficiency while preserving cell viability. Using STED microscopy, we have shown that the cell membrane opens transiently and locally on top of the straws when nanostraw-electroporation is perfomed.
More recently, we utilized GFP-encoding DNA plasmids stained with an intercalating dye for immediate analysis of the electrophoretic transport of the cargo, and assessed GFP expression 48 h later for transfection efficiency. We compared the transfection efficiency between nanostraws and nanopores. While the amount of GFP plasmids delivered using nanostraws was greater than that delivered through nanopores, the resulting GFP fluorescence measured 48 hours post-transfection was lower for the nanostraws. Moreover, fewer cells were observed on the substrate 48 hours after transfection with nanostraws compared to those using nanopores. This suggests that nanopore electroporation, being a simpler technique, presents an appealing alternative to nanostraws for achieving effective and gentle cell transfection.