Stimulating Cellular Appetite: Electro-Mechanical Poration Boosts Lipid Nanoparticle-Mediated Gene Delivery in Blood Cells

We present an unprecedented transfection method utilizing electro-mechanical-poration (EMP) in combination with lipid nanoparticles (LNPs). In contrast to conventional non-viral transfection methods-including electroporation (EP), mechanoporation, and passive LNPmediated delivery-our approach actively stimulates cellular uptake of LNPs through localized electric fields and microbubble formation. We prepared three types of nanometer-sized artifacts (nano-artifacts) for transfection: bare plasmid DNA (pDNA), pDNA-cationic polymer complex (polyplex), and LNP containing pDNA. These nano-artifacts were applied to blood cells with or without EP or EMP. We achieved the highest transfection efficiency of blood cells by using LNP combined with EMP, with stable gene expression observed for at least one week. These findings suggest that EMP enables effective delivery of nano-artifacts by enhancing cellular appetite, while EP increased membrane permeability by creating pores in the cell membrane. Therefore, the proposed method will contribute to advancements in cell engineering and therapeutic applications.