Palmitic acid-modified poly-L-lysine for non-viral delivery of plasmid DNA to skin fibroblasts.
Palmitic acid conjugates of poly-L-lysine (PLL-PA) were prepared, and their ability to deliver plasmid DNA into human skin fibroblasts was evaluated in vitro. The conjugates were capable of condensing a 4.7 kb plasmid DNA into 50-200 nm particles (mean +/- SD = 112 +/- 34 nm), which were slightly smaller than the particles formed by PLL (mean +/- SD = 126 +/- 51 nm). Both PLL and PLL-PA were readily taken up by the cells, but PLL-PA delivered the plasmid DNA into a higher proportion of cells. DNA delivery was found to be reduced by endocytosis inhibitor Brefeldin A, suggesting an active mechanism of particle uptake. Using enhanced green fluorescent protein (EGFP) as a reporter gene, PLL-PA was found to give the highest number of EGFP-positive cells among several carriers tested, including polyethyleneimine, Lipofectamine-2000, and an adenovirus. Although some carriers gave a higher percentage of EGFP-positive cells than PLL-PA, they were also associated with higher toxicities. We conclude that PLL-PA is a promising gene carrier for non-viral modification of human fibroblasts.
Abbasi
Meysam
M
Department of Biomedical Engineering, University of Alberta, Edmonton, AB, Canada.
Uludag
Hasan
H
Incani
Vanessa
V
Olson
Cori
C
Lin
Xiaoyue
X
Clements
Başak Açan
BA
Rutkowski
Dorothy
D
Ghahary
Aziz
A
Weinfeld
Michael
M
D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics*; D004337:Drug Carriers / Q000138:chemical synthesis / Q000737:chemistry* / Q000493:pharmacokinetics; D005347:Fibroblasts / Q000737:chemistry / Q000378:metabolism*; D018014:Gene Transfer Techniques; D006801:Humans; D015394:Molecular Structure; D019308:Palmitic Acid / Q000737:chemistry* / Q000378:metabolism; D010316:Particle Size; D010957:Plasmids / Q000235:genetics; D011107:Polylysine / Q000737:chemistry* / Q000378:metabolism; D012867:Skin / Q000378:metabolism; D013997:Time Factors
D004337:Drug Carriers; D011107:Polylysine; D019308:Palmitic Acid; D004247:DNA