Mechanism of jet injector-induced plasmid DNA uptake: Contribution of shear stress and endocytosis

Abstract The administration of plasmid DNA (pDNA) using a pyro-drive jet injector allows gene expression in cells of the treated tissue; however, the detailed plasmid uptake process remains to be determined. A recent theory suggests that shear stress enhances the endocytosis pathway and pDNA internalization. Here, we investigated the process of pDNA uptake in the context of a pyro-drive jet injector-based administration as a way to optimize gene transfer efficiency via the increase in DNA uptake. The gene expression was significantly improved when the shear stress caused by the jet was generated where the pDNA was retained. Contrarily, heparin, an inhibitor of the spontaneous uptake of injected DNA, inhibited the gene expression in jet injection. In addition, treatment with typical endocytosis inhibitors (chlorpromazine, methyl-β-cyclodextrin, dimethyl amiloride, rottlerin, and NSC23766) also reduced plasmid expression efficiency in the context of jet injection; conversely, endosome escape in the context of chloroquine treatment increased the expression efficiency. Altogether, our results not only clarify the mechanism of pDNA uptake in the context of jet injection but also highlight the key role of endosomes on the intracellular trafficking of pDNA. Importantly, such findings may impact other studies on gene transfer and endocytosis and boost further efforts to improve the efficiency and safety of jet injection in the context of both basic and translational applications.