In vivo time-dependent gene expression of cationic lipid-based emulsion as a stable and biocompatible non-viral gene carrier.
To make stable and biocompatible non-viral gene carriers for therapeutic gene therapy, we developed a cationic lipid-based emulsion (CLE) prepared by an oil-in-water (O/W) emulsion method, wherein squalene oil was used as an oil core and the cationic lipid, 1,2-dioleoyl-sn-glycero-3-trimethylammonium-propane (DOTAP), was employed as an emulsifier. To evaluate in vivo characteristics such as toxicity and time-dependent gene expression, a bioluminescence reporter gene in pCMV-luc plasmid DNA was simply mixed with CLE in aqueous condition, resulting in a CLE/DNA complex. The CLE/DNA complex was optimized to form a compact and stable nano-sized particle by adding different amounts of plasmid DNA, and an optimal cationic lipid-to-DNA (C/D) weight ratio of 4 was identified. Freshly prepared CLE/DNA complex, with a C/D of 4, showed a high transfection efficiency and minimal cytotoxicity in vitro, compared to controls of a liposome (DOTAP)/DNA complex and a branched poly(ethyleneimine) (Mw=25 kDa) (bPEI)/DNA complex, respectively. The in vivo characteristics of the CLE/DNA complex were evaluated after intravenous injection into Balb/c mice. Time-dependent gene expression data in vivo were obtained using a non-invasive, whole animal bioluminescence imaging system. These data showed that the CLE/DNA complex offered prolonged high-level gene expression for 1 week, particularly in the liver and spleen. On the other hand, the controls of DOTAP/DNA complex and bPEI/DNA complex showed a relatively lower gene expression, because of the unstable and toxic properties of the control carriers. Our in vivo gene expression data demonstrate the potential of the CLE/DNA complex as a non-viral gene carrier for in vivo gene delivery.
Kwon
Seok Min
SM
Biomedical Research Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seongbuk-gu, Seoul 136-791, South Korea.
Nam
Hae Yun
HY
Nam
Taehwan
T
Park
Kyeongsoon
K
Lee
Seulki
S
Kim
Kwangmeyung
K
Kwon
Ick Chan
IC
Kim
Joon
J
Kim
Jun
J
Kang
Dongmin
D
Park
Jae Hyung
JH
Jeong
Seo Young
SY
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics*; D045703:Emulsifying Agents / Q000737:chemistry / Q000493:pharmacokinetics*; D004655:Emulsions; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry / Q000493:pharmacokinetics*; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors; D008156:Luciferases / Q000235:genetics*; D049449:Luminescence; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000737:chemistry; D010957:Plasmids / Q000235:genetics; D000644:Quaternary Ammonium Compounds / Q000737:chemistry / Q000493:pharmacokinetics*; D013185:Squalene / Q000737:chemistry / Q000493:pharmacokinetics*; D014018:Tissue Distribution
D045703:Emulsifying Agents; D004655:Emulsions; D005229:Fatty Acids, Monounsaturated; D000644:Quaternary Ammonium Compounds; D013185:Squalene; D004247:DNA; D008156:Luciferases; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane