Intracellular siRNA delivery system using polyelectrolyte complex micelles prepared from VEGF siRNA-PEG conjugate and cationic fusogenic peptide.
To develop a small interfering RNA (siRNA) delivery system with low cytotoxicity and high transfection efficiency, siRNA was conjugated to poly(ethylene glycol) via a disulfide linkage (siRNA-PEG) to prepare polyelectrolyte complex micelles (PECMs) by condensing with a cationic fusogenic peptide (KALA). The siRNA-PEG conjugate exhibited enhanced resistance to degradation from nucleases. Anionic siRNA-PEG conjugate and cationic KALA, when mixed in an aqueous phase, spontaneously formed nano-sized PECMs (<200nm) that have an inner core of charge neutralized siRNA/KALA complex surrounded by a PEG corona. Vascular endothelial growth factor (VEGF) siRNA was used to demonstrate VEGF sequence-specific gene inhibition in prostate carcinoma cells (PC-3 cells). The extent of gene silencing was gradually increased with increasing nitrogen to phosphate (N/P) ratio and the concentration of siRNA-PEG/KALA PECMs. These results suggest that the formulation of siRNA-PEG/KALA PECMs could be widely applied for intracellular delivery of various therapeutic siRNAs.
Lee
Soo Hyeon
SH
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, South Korea.
Kim
Sun Hwa
SH
Park
Tae Gwan
TG
D001672:Biocompatible Materials / Q000737:chemistry; D045744:Cell Line, Tumor; D016503:Drug Delivery Systems / Q000379:methods*; D004573:Electrolytes / Q000737:chemistry; D020868:Gene Silencing; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008297:Male; D008823:Micelles; D011092:Polyethylene Glycols / Q000737:chemistry*; D011471:Prostatic Neoplasms / Q000503:physiopathology*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
D001672:Biocompatible Materials; D004573:Electrolytes; D008081:Liposomes; D008823:Micelles; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; D011092:Polyethylene Glycols