The first step toward hydrophobic polymer-based nanospheres for gene delivery is to encapsulate and release plasmid DNA. However, encapsulating large hydrophilic molecules in very small nanospheres has been difficult, and only a few examples exist in the literature. For example, maximizing protein and peptide as well as small molecule encapsulation requires adjustments in pH or addition of excipients to charge neutralize, and make less hydrophilic, the compound to be encapsulated. Following this model, we have used a cationic lipid to load and release plasmid DNA from nanospheres made by the phase inversion/solvent diffusion method.