Transfection with Nanostructure Electro-Injection is Minimally Perturbative

Abstract

Transfection is a critical step for gene editing and cell-based therapies. Nanoscale technologies have shown great promise in providing higher transfection efficiency and lower cell perturbation than conventional viral, biochemical, and electroporation techniques due to their small size and localized effects. Although this has significant implications for using cells post-transfection, it has not been thoroughly studied. Here, the nano-electro-injection (NEI) platform is developed, which makes use of localized electric fields to transiently open pores on cell membranes followed by electrophoretic delivery of DNA into cells. NEI provides twofold higher net transfection efficiency than biochemicals and electroporation in Jurkat cells. Analysis of cell doubling time, intracellular calcium levels, and messenger ribonucleic acids (mRNA) expression changes after these gene delivery methods reveals that viruses and electroporation adversely affected cell behavior. Cell doubling times increase by more than 40% using virus and electroporation methods indicative of higher levels of cell stress, unlike NEI which only minimally affects cell division. Finally, electroporation, but not NEI, greatly alters the expression of immune-associated genes related to immune cell activation and trafficking. These results highlight that nanoscale delivery tools can have significant advantages from a cell health perspective for cell-based research and therapeutic applications.