Evaluation and mechanism studies of PEGylated dendrigraft poly-L-lysines as novel gene delivery vectors.
Dendrimers have attracted great interest in the field of gene delivery due to their synthetic controllability and excellent gene transfection efficiency. In this work, dendrigraft poly-L-lysines (DGLs) were evaluated as a novel gene vector for the first time. Derivatives of DGLs (generation 2 and 3) with different extents of PEGylation were successfully synthesized and used to compact pDNA as complexes. The result of gel retardation assay showed that pDNA could be effectively packed by all the vectors at a DGLs to pDNA weight ratio greater than 2. An increase in the PEGylation extent of vectors resulted in a decrease in the incorporation efficiency and cytotoxicity of complexes in 293 cells, which also decreased the zeta potential a little but did not affect the mean diameter of complexes. Higher generation of DGLs could mediate higher gene transfection in vitro. Confocal microscopy and cellular uptake inhibition studies demonstrated that caveolae-mediated process and macropinocytosis were involved in the cellular uptake of DGLs-based complexes. Also the results indicate that proper PEGylated DGLs could mediate efficient gene transfection, showing their potential as an alternate biodegradable vector in the field of nonviral gene delivery.
Department of Pharmaceutics, School of Pharmacy, Fudan University, Shanghai 201203, People's Republic of China.
Rongqin
R
Huang
Shuhuan
S
Liu
Kun
K
Shao
Liang
L
Han
Weilun
W
Ke
Yang
Y
Liu
Jianfeng
J
Li
Shixian
S
Huang
Chen
C
Jiang
D016923:Cell Death; D002460:Cell Line; D002470:Cell Survival; D004247:DNA / Q000378:metabolism; D050091:Dendrimers / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010316:Particle Size; D011092:Polyethylene Glycols / Q000737:chemistry*; D011107:Polylysine / Q000031:analogs & derivatives* / Q000737:chemistry*; D014162:Transfection
D050091:Dendrimers; D049452:Green Fluorescent Proteins; D011107:Polylysine; D011092:Polyethylene Glycols; D004247:DNA; D008156:Luciferases