Abstract
Primary human fibroblasts and a series of cell lines (A549, BNL CL.2, H225, NIH 3T3 and Rat-1) are efficiently transfected by using positively charged complexes of plasmid DNA and transferrin-polylysine or polylysine in the presence of glycerol (1 molar to 1.8 molar, depending on the cell type). An increase in gene expression of up to several-hundredfold (compared to complexes without glycerol) is obtained if the transfection mixture is incubated with the cells for 3-4 h at 37 degrees C. This simple method has been used for transient expression of luciferase, beta-galactosidase and interleukin-2, and also for the generation of stably transfected cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Anti-Bacterial Agents / pharmacology
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Cell Adhesion / drug effects
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Cell Line
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Chloroquine / pharmacology
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Clone Cells / drug effects
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DNA / chemistry
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Enzyme Inhibitors / pharmacology
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Ethylene Glycol
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Ethylene Glycols / pharmacology
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Fibroblasts / cytology
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Gene Expression Regulation
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Genes, Reporter
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Glycerol / chemistry*
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Humans
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Interleukin-2 / biosynthesis
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Ligands*
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Macrolides*
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Plasmids
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Polylysine / chemistry*
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Promoter Regions, Genetic
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Transfection / methods*
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beta-Galactosidase / analysis
Substances
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Anti-Bacterial Agents
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Enzyme Inhibitors
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Ethylene Glycols
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Interleukin-2
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Ligands
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Macrolides
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Polylysine
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Chloroquine
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bafilomycin A1
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DNA
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beta-Galactosidase
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Ethylene Glycol
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Glycerol