Calcium condensation of DNA complexed with cell-penetrating peptides offers efficient, noncytotoxic gene delivery.

Drug delivery strategies using cell-penetrating peptides (CPPs) have been widely explored to improve the intracellular delivery of a large number of cargo molecules. Electrostatic complexation of plasmid DNA using CPPs has been less explored due to the relatively large complexes formed and the low levels of gene expression achieved when using these low-molecular-weight polycations as DNA condensing agents. Here, condensing nascent CPP polyplexes using CaCl(2) produced small and stable nanoparticles leading to gene expression levels higher than observed for control polyethylenimine gene vectors. This simple formulation approach showed negligible cytotoxicity in A549 lung epithelial cells and maintained particle size and transfection efficiency even in the presence of serum.