Reducible cationic lipids for gene transfer

被引:68
作者
Wetzer, B
Byk, G
Frederic, M
Airiau, M
Blanche, F
Pitard, B
Scherman, D
机构
[1] Ecole Natl Super Chim Paris, Lab Chim Bioorgan & Biotechnol Mol & Cellulaire, UMR 7001, CNRS,Aventis Pharma,Ctr Rech Vitry Alfortville, F-94403 Vitry, France
[2] Bar Ilan Univ, Dept Chem, Lab Peptidomimet & Genet Chem, IL-52900 Ramat Gan, Israel
[3] Rhodia, CRA, F-93308 Aubervilliers, France
[4] Aventis Pasteur, F-69280 Marcy Etoile, France
关键词
cationic lipid transfection; disulphide-bond-bearing lipid; gene therapy; non-viral gene transfer; reducible DNA-lipid complexes;
D O I
10.1042/0264-6021:3560747
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
One of the main challenges of gene therapy remains the increase of gene delivery into eukaryotic cells. We tested whether intracellular DNA release, an essential step for gene transfer, could be facilitated by using reducible cationic DNA-delivery vectors. For this purpose, plasmid DNA was complexed with cationic lipids bearing a disulphide bond, This reduction-sensitive linker is expected to be reduced and cleaved in the reducing milieu of the cytoplasm, thus potentially improving DNA release and consequently transfection, The DNA-disulphide-lipid complexation was monitored by ethidium bromide exclusion, and the size of complexes was determined by dynamic light scattering, It was found that the reduction kinetics of disulphide groups in DNA-lipid complexes depended on the position of the disulphide linker within the lipid molecule, Furthermore, the internal structure of DNA-lipid particles was examined by small-angle X-ray scattering before and after lipid reduction. DNA release from lipid complexes was observed after the reduction of disulphide bonds of several lipids. Cell-transfection experiments suggested that complexes formed with selected reducible lipids resulted in up to 1000-fold higher reporter-gene activity, when compared with their analogues without disulphide bonds. In conclusion, reduction-sensitive groups introduced into cationic lipid backbones potentially allow enhanced DNA release from DNA-lipid complexes after intracellular reduction and represent a tool for improved vectorization.
引用
收藏
页码:747 / 756
页数:10
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