Glycerol enhancement of ligand-polylysine/DNA transfection.
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.
Zauner
W
W
Research Institute of Molecular Pathology, University of Vienna, Austria.
Kichler
A
A
Schmidt
W
W
Sinski
A
A
Wagner
E
E
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D002999:Clone Cells / Q000187:drug effects; D004247:DNA / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019855:Ethylene Glycol; D005026:Ethylene Glycols / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005990:Glycerol / Q000737:chemistry*; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis; D008024:Ligands; D018942:Macrolides; D010957:Plasmids; D011107:Polylysine / Q000737:chemistry*; D011401:Promoter Regions, Genetic; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000032:analysis
D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D005026:Ethylene Glycols; D007376:Interleukin-2; D008024:Ligands; D018942:Macrolides; D011107:Polylysine; D002738:Chloroquine; C040929:bafilomycin A1; D004247:DNA; D001616:beta-Galactosidase; D019855:Ethylene Glycol; D005990:Glycerol