[Preparation of lipid-protamine-DNA complexes and evaluation of their transfection efficiencies in vitro].
OBJECTIVE
To develop a novel non-viral gene delivery systems lipid-polycation-DNA complexes (LPD) and investigate their transfection efficiencies in vitro.


METHODS
LPD were prepared as follows by first mixing the plasmid DNA and protamine together, then the resulted polyplexes were incubated for 10 min at room temperature, followed by addition of preformed cationic liposomes. The morphology of LPD was observed by transmission electron microscopy. The diameter and surface charge of LPD were measured by photon correlation spectroscopy (PCS). The nuclease protection ability of LPD was evaluated by agarose gel electrophoresis. Estimation of transfection efficiency was performed by galactosidase assay in Chang, HepG2 and SMMC-7721 cells.


RESULTS
The average diameter and the zeta potential of LPD were 143.5 nm and 32.6 mV, respectively. LPD could protect the plasmid DNA from nuclease degradation after 2 hours incubation at 37 degrees C while the naked DNA degraded rapidly. The average transfection efficiencies were (69 +/- 6)%, (43 +/- 7)% and (96.2 +/- 1.8)% in Chang cells, HepG2 cells and SMMC-7721 cells respectively.


CONCLUSIONS
LPD could be prepared easily with small particle sizes and high transfection activities. LPD may be good non-viral vectors for applications in gene delivery.
Sun
Xun
X
West China School of Pharmacy, Sichuan University, Chengdu 610041, China.
Zhang
Zhi-rong
ZR
D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004247:DNA; D005822:Genetic Vectors; D022781:Hepatocytes / Q000166:cytology; D006801:Humans; D008081:Liposomes; D008113:Liver Neoplasms / Q000473:pathology; D010316:Particle Size; D011479:Protamines; D014162:Transfection
D008081:Liposomes; D011479:Protamines; D004247:DNA