Quantitative aspects of endocytic activity in lipid-mediated transfections

Variation in transfection efficiency observed in different cell-types is poorly understood. To investigate the influence of endocytic activity on lipid-mediated transfections, we have monitored both the processes in 12 different cell-types. The endocytic activity shows a strong positive correlation...

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Bibliographic Details
Published in:FEBS letters 2005-05, Vol.579 (12), p.2635-2642
Main Authors: Prasad, Tekkatte Krishnamurthy, Rangaraj, Nandini, Rao, Nalam Madhusudhana
Format: Article
Language:English
Subjects:
1,2-dioleoyl-3-trimethylammonium-propane
Androstadienes - pharmacology
Animals
Bisbenzimidazole
Cationic lipid
cationic lipid DNA complexes
Cell Division
Cell Line
Cell Line, Tumor
Cercopithecus aethiops
CHO Cells
CLDC
COS Cells
Cricetinae
Cricetulus
dimethyl sulfoxide
DMEM
DMSO
DOPE
DOTAP
Dulbecco's modified Eagle's medium
Endocytosis
Endocytosis - drug effects
Endocytosis - genetics
Enzyme Inhibitors - pharmacology
Fatty Acids, Monounsaturated - metabolism
FITC
fluorescein isothiocyanate
Fluorescein-5-isothiocyanate
Fluorescent Dyes
Gene delivery
Genes, Reporter
HeLa Cells
horseradish peroxidase
HRP
Humans
L Cells (Cell Line)
l-α-dioleoyl phosphatidylethanolamine
Liposomes
Mice
Microscopy, Confocal
Mitosis
N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycerol-3-phosphaethanolamine
NBD-PE
NF-kappa B - analysis
NF-kappa B - metabolism
NIH 3T3 Cells
PBS
phosphate buffered saline
Phosphatidylethanolamines - metabolism
Phosphatidylinositol 3-Kinases - antagonists & inhibitors
Quaternary Ammonium Compounds - metabolism
Rh-DHPE
rhodamine B 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine
Rhodamines
Transfection
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Summary:Variation in transfection efficiency observed in different cell-types is poorly understood. To investigate the influence of endocytic activity on lipid-mediated transfections, we have monitored both the processes in 12 different cell-types. The endocytic activity shows a strong positive correlation ( P < 0.01), with transfection efficiency. Treatment with wortmannin resulted in cell-type-dependent inhibition of transfection. Studies on M-phase cells by confocal microscopy show that compared to interphase cells, uptake of cationic liposomes was substantially reduced. In addition, transfection efficiency of cells in mitotic phase was inhibited by >70% compared to controls. Our study based on several cell-types demonstrates for the first time that quantitative aspects of endocytosis have decisive influence on the overall process of lipid-mediated transgene expression.
ISSN:0014-5793
1873-3468
DOI:10.1016/j.febslet.2005.03.083