The construction of a novel kind of non-viral gene delivery vector based on protein as core backbone.
OBJECTIVE
A novel kind of non-viral gene delivery vector based on transferrin (Tf) as the core component was constructed with high transfection efficiency and low toxicity.


METHODS
The synthesis vector of Tf-PEI600 was confirmed by different physicochemical methods, including (1)H nuclear magnetic resonance, gel permeation chromatography, X-ray and thermogravimetric analysis. The cytotoxicity and gene delivery efficiency of the synthesized vector were verified by in vitro experiments.


RESULTS
The agarose gel electrophoresis assay indicated that the novel copolymer Tf-PEI600 could efficiently condense plasmid DNA and the condensed nanoparticles exhibited a spherical shape. As the weight ratio of Tf-PEI600 to DNA reached 15.0, the particle size (about 200 nm) and the zeta potential (about 20 mV) of the nanoparticles became optimal for gene delivery. The methylthiazolyl tetrazolium (MTT) assay showed the cytotoxicity of Tf-PEI600 to be similar to that of PEI600 and much lower than that of PEI25kDa. In gene-delivery experiments with COS-7 cells and HepG2 cells, the Tf-PEI600 showed about a 30- to 53-fold higher efficiency than PEI600 and nearly equal to that of PEI25kDa.


CONCLUSIONS
These data suggest that Tf-PEI600, with the advantages of low toxicity and high gene-delivery efficiency, might have great prospects in the practice of gene delivery. The core-shell structure of Tf-PEI600 also provided a novel strategy for the construction of non-viral gene delivery vectors.
Li
D
D
Department of Oncology, Sir Run Run Shaw Hospital affiliated to School of Medicine, Zhejiang University, Hangzhou 310003, China.
Kong
Y
Y
Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China.
Yu
H
H
Institute of Immunology, Zhejiang University, Hangzhou 310031, China.
Lehtinen
A
A
Department of Chemical Technology, Helsinki University of Technology, FIN-02015 TKK, Finland.
Huang
H
H
Institute of Immunology, Zhejiang University, Hangzhou 310031, China.
Shen
F
F
Institute of Immunology, Zhejiang University, Hangzhou 310031, China.
Min
L
L
Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China.
Zhou
J
J
Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China.
Tang
G
G
Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China.
Wang
Q
Q
Institute of Immunology, Zhejiang University, Hangzhou 310031, China.
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D002850:Chromatography, Gel; D004274:DNA, Recombinant / Q000737:chemistry / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D046529:Microscopy, Electron, Transmission; D008970:Molecular Weight; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity / Q000648:ultrastructure; D010316:Particle Size; D011094:Polyethyleneimine / Q000737:chemistry / Q000633:toxicity; D014162:Transfection; D014168:Transferrin / Q000737:chemistry* / Q000633:toxicity
D004274:DNA, Recombinant; D014168:Transferrin; D011094:Polyethyleneimine