Novel Fatty Acid Modifications of Transportan 10

Cell-penetrating peptides (CPPs) are able to efficiently internalize into cells and can therefore be used as vectors for non-viral cellular delivery of different cargoes. Previous studies have shown that hydrophobic modifications of different CPPs can increase their transfection efficiency dramatically. In this study we have modified the cell penetrating-peptide transportan 10 (TP10) with a variety of hydrophobic molecules to determine the role of hydrophobicity in the uptake of these molecules. The results can be used to synthesize more efficient delivery vectors. To evaluate how these constructs are able to transport cargoes into cells we used 2′-OMe splice correcting oligonucleotides. Non-covalent peptide-cargo complexes were formed and their transfection efficiency was measured using a luciferase readout system. The hydrophobicity of the novel modifications was correlated with their biological efficacy. We determined the most efficient range of hydrophobicity for TP10 analogs for delivering oligonucleotides into cells. In order to assess how the transfection efficacy of these particles is dependent on their size the hydrodynamic diameter of the formed nanoparticles was measured using dynamic light scattering. These findings will be used to develop highly efficient non-viral gene therapy vectors.