title: Optimal lipofection reagent varies with the molecular modifications of the DNA.
abstract: Cationic lipid reagents differ in their cytofection efficacy with different cell types. No evidence has addressed whether the same lipid reagent is best for different DNAs in a single cell line. Immortalized avian embryonic cardiomyocytes cultured in vitro were tested with 15 cationic lipid reagents using (A) a beta-gal expression plasmid, (B) a fluorescein-tagged, phosphorothioate-modified ODN B, (C) a fluorescein-tagged, ethoxy-modified ODN C with the same nucleotide sequence as ODN B, and (D) a fluorescein-tagged, phosphorothioate-modified ODN D with a different nucleotide sequence from ODNs B and C. Cytofection was scored as percent of cells expressing beta-gal activity or showing diffuse cellular fluorescence. The best lipid reagents for the phosphorothioate-modified ODNs were ODN-specific and markedly different from the best lipid reagents for the expression plasmid or for the ethoxy-modified ODN. These results suggest that the best cationic lipid reagent for a particular cell type varies with the physical and chemical form of the DNA being transfected into the cells.
authors.affiliation: Division of Biology, Kansas State University, Manhattan 66506-4901, USA.
authors.forename: A H
authors.initials: AH
authors.lastname: Conrad
authors.forename: M A
authors.initials: MA
authors.lastname: Behlke
authors.forename: T
authors.initials: T
authors.lastname: Jaffredo
authors.forename: G W
authors.initials: GW
authors.lastname: Conrad
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D008055:Lipids / Q000494:pharmacology; D009206:Myocardium / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011784:Quail; D014162:Transfection / Q000187:drug effects; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D008055:Lipids; D004247:DNA; D001616:beta-Galactosidase