Rational Design and Assessment of Protein-Lipid Hybrid Vehicle for Non-Viral Gene Delivery

Gene therapy has the potential to treat various diseases, but a vehicle capable of safely delivering nucleic acids does not currently exist for widespread clinical use. We have recently developed a lipoproteoplex (LPP) consisting of a super-charged coiled-coil protein (CSP) and a cationic liposomal carrier, that has the ability to condense nucleic acids and deliver them in vivo. A CSP library has been rationally designed to improve the efficacy of the LPP compared to the parent protein via increased nucleic acid binding and endosomal escape. The secondary structure and nucleic acid binding ability of each library member was assessed using circular dichroism and electrophoretic mobility shift assay, then compared to functional transfection data. Structural and functional data suggests that both high alpha-helicity of the protein component of the LPP as well as high nucleic acid binding affinity are crucial for efficient gene delivery into cells.