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Enhanced in vitro and in vivo cationic lipid‐mediated gene delivery with a fluorinated glycerophosphoethanolamine helper lipid
Author(s) -
Boussif Otmane,
Gaucheron Jérôme,
Boulanger Caroline,
Santaella Catherine,
Kolbe Hanno V. J.,
Vierling Pierre
Publication year - 2001
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.166
Subject(s) - luciferase , transfection , in vivo , in vitro , microbiology and biotechnology , reporter gene , genetic enhancement , a549 cell , chemistry , gene delivery , biology , biochemistry , gene expression , gene
Abstract Background One of the main drawbacks of synthetic, non‐viral gene vectors is their relatively low in vivo efficiency when compared with viral vectors. The present paper describes the use of a partially fluorinated glycerophosphoethanolamine ( F ‐PE), a close analog of DOPE, which, as a helper lipid with the cationic lipopolyamine pcTG90, increases its in vitro and in vivo gene transfer capability to a larger extent than DOPE. Methods To evaluate the contribution of F ‐PE to lipoplex‐mediated gene transfer, the effect of including F ‐PE in lipoplexes formulated with the lipopolyamine pcTG90 for various pcTG90/DOPE/ F ‐PE molar ratios [1:(1−x):x; 1:(2−y):y] was examined. For the in vitro analyses on human lung carcinoma epithelial A549 cells, the lipoplexes were formulated with the luciferase reporter plasmid pTG11033 using various N/P ratios (from 10 to 0.8, N=number of pcTG90 amines, P=number of DNA phosphates). The in vivo analyses were performed (1) with the luciferase reporter plasmid pCMV‐Luc, which gives higher luciferase expression in the lung than pcTG11033; (2) with pcTG90/co‐lipid(s) (1:2) lipoplexes which yield higher expression than the (1:1) formulations; and (3) by intravenous (iv) injection into the tail vein of mice. Results The efficiency of the F ‐PE lipoplexes to transfect in vitro A549 cells was significantly higher (5–90‐fold) than that of DOPE lipoplexes, when formulated in HEPES. However, when formulated in 5% glucose, both co‐lipids display a comparable transfection helper potential. Most remarkably, an up to eight‐fold increase of luciferase expression could be measured in the lung after iv injection of pcTG90/ F ‐PE (1:2) N/P 5 lipoplexes as compared with the pcTG90/DOPE lipoplexes. It led also to higher luciferase expression than PEI(ExGen500)/pCMV‐Luc N/P 10 polyplexes. Besides expression in lung, low levels of luciferase expression were also observed in heart, spleen and liver. Conclusion The present work, showing a higher in vitro and in vivo transfection potential for lipoplexes formulated with a partially fluorinated co‐lipid as compared with its analogous DOPE lipoplexes or PEI polyplexes, indicates that ‘fluorinated’ lipoplexes are attractive candidates for in vivo applications. Copyright © 2001 John Wiley & Sons, Ltd.