Chemically Tuned Intracellular Gene Delivery by Core‐Shell Nanoparticles: Effects of Proton Buffering, Acid Degradability, and Membrane Disruption

Nanoparticles consisting of a condensed nucleic acid core surrounded by protective layers which aid to overcome extracellular and intracellular hurdles to gene delivery (i. e., core‐shell nanoparticles, CSNPs) synthetically mimic viruses. The outer shells shield the core and are particularly designed to enable facilitated release of the gene payload into the cytoplasm, the major limiting step in intracellular gene delivery. The hypothetical proton sponge effect and degradability in response to a stimulus (i. e., mildly acidic pH in the endosome) are two prevailing, although contested, principles in designing effective carriers for intracellular gene delivery via endosomal escape. Utilizing the highly flexible chemical‐tuning of the polymeric shell via surface‐initiated photo‐polymerization of the various monomers at different molecular ratios, the effects of proton buffering capacity, acid‐degradability, and endosomal membrane‐lysis property on intracellular delivery of plasmid DNA by CSNPs were investigated. This study demonstrated the equivalently critical roles of proton buffering and acid‐degradability in achieving efficient intracellular gene delivery, independent of cellular uptake. Extended proton buffering resulted in further improved transfection as long as the core structure was not compromised. The results of the study present a promising synthetic strategy to the development of an efficient, chemically‐tunable gene delivery carrier. Core‐shell nanoparticles (CSNPs) with a pDNA/protamine sulfate polyplex core and a polymeric shell were prepared by polymerizing various monomers and crosslinkers that are designed to withstand the intracellular environment. Endosomal escape of CSNPs was found to be cooperatively facilitated by proton buffering, acid‐degradability, and moderate membrane disruption.