Engineering Endosomal Escape: Improving mRNA Delivery Using Novel Lipid Micelle Systems

This study evaluated mRNA delivery efficiency in PC3 prostate cancer cells using micelle-based lipid nanoparticles (DOTAP-, DOPE-, DOPC-, and MVL5-containing formulations) compared to Lipofectamine as a positive control. Lipid films were prepared, hydrated, and complexed with EGFP mRNA before dosing cells under controlled serum and buffer conditions. Live-cell fluorescence microscopy assessed uptake, lipid distribution, and GFP translation. DOTAP/DOPE formulations produced the most consistent transfection, while MVL5-based complexes showed strong uptake but no cytoplasmic release, suggesting an endosomal escape barrier. These results highlight how lipid composition, charge ratio, and complexation environment critically shape mRNA delivery outcomes.