IONP-PLL: a novel non-viral vector for efficient gene delivery.
BACKGROUND
Non-viral methods of gene delivery have been an attractive alternative to virus-based gene therapy. However, the vectors that are currently available have drawbacks limiting their therapeutic application.


METHODS
We have developed a self-assembled non-viral gene carrier, poly-L-lysine modified iron oxide nanoparticles (IONP-PLL), which is formed by modifying poly-L-lysine to the surface of iron oxide nanoparticles. The ability of IONP-PLL to bind DNA was determined by ratio-dependent retardation of DNA in the agarose gel and co-sedimentation assay. In vitro cytotoxic effects were quantified by MTT assay. The transfection efficiency in vitro was evaluated by delivering exogenous DNA to different cell lines using IONP-PLL. Intravenous injection of IONP-PLL/DNA complexes into mice was evaluated as a gene delivery system for gene therapy. The PGL2-control gene encoding firefly luciferase and the EGFP-C2 gene encoding green fluorescent protein were used as marker genes.


RESULTS
IONP-PLL could bind and protect DNA. In contrast to PLL and cationic liposomes, IONP-PLL described here was less cytotoxic in a broad range of concentrations. In the current study, we have demonstrated that IONP-PLL can deliver exogenous gene to cells in vitro and in vivo. After intravenous injection, IONP-PLL transferred reporter gene EGFP-C2 to lung, brain, spleen and kidney. Furthermore, we have demonstrated that IONP-PLL transferred exogenous DNA across the blood-brain barrier to the glial cells and neuron of brain.


CONCLUSIONS
IONP-PLL, a low-toxicity vector, appears to have potential for fundamental research and genetic therapy in vitro and in vivo, especially for gene therapy of CNS disease.
Xiang
Juan-Juan
JJ
Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, Hunan, China.
Tang
Jing-Qun
JQ
Zhu
Shi-Guo
SG
Nie
Xin-Min
XM
Lu
Hong-Bin
HB
Shen
Shou-Rong
SR
Li
Xiao-Ling
XL
Tang
Ke
K
Zhou
Ming
M
Li
Gui-Yuan
GY
D000818:Animals; D001921:Brain / Q000648:ultrastructure; D002460:Cell Line; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D005290:Ferric Compounds / Q000737:chemistry* / Q000378:metabolism; D005347:Fibroblasts / Q000737:chemistry; D018390:Gene Targeting; D005822:Genetic Vectors / Q000008:administration & dosage / Q000633:toxicity; D049452:Green Fluorescent Proteins; D006801:Humans; D007668:Kidney / Q000648:ultrastructure; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009457:Neuroglia / Q000737:chemistry; D011107:Polylysine / Q000008:administration & dosage / Q000378:metabolism*; D014018:Tissue Distribution; D014162:Transfection / Q000379:methods*
D005290:Ferric Compounds; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; C000499:ferric oxide; D011107:Polylysine; D004247:DNA