초록 |
Plasmid DNA-cationic lipid complexes have a limiting transfection such as distribution by dnase and toxic side effect both in vitro and in vivo. In this research, we report the ‘stabilized plasmid-lipid particles’ (SPLP) using enzymatically cleavable oligo-peptide. Particle makes the detergent dialysis method and can be encapsulation plasmid DNA. Also, the SPLP composed of DOPE, the cationic lipid DOTAP and PEG5000-GLFG-K-C16. GFLG (Gly-Phe-Leu-Gly) sequence is known to be cleaved by enzyme Cathepsin-B and may promote endosomal escape after intracellular internalization. We purify the non-encapsulation DNA by DEAE-separose chromatography and measure size diameter by DLS. The encapsulation DNA is analyzed by lysis to TritionX-100 and Pico-Green assay. Furthermore, the transfection efficiency and cytotoxicity were determined by luciferase assay and MTT assay. |