Novel cationic liposomes for DNA-transfection with high efficiency and low toxicity.
Liposomes containing the natural cationic amphiphile, sphingosine and some of its derivatives were used for transfection of DNA in vitro. Multilamellar liposomes comprised of dioleoylphosphatidylethanolamine (DOPE), different sphingosine derivatives, and diacylglycerols with varying fatty acid chains, preincubated with DNA, transfected efficiently the KK-1 murine granulosa cells. Most efficient transfection on this cell line was achieved with liposomes composed of phytosphingosine, DOPE, and dioctanoylglycerol (DC8G) (64:31:4.8, molar stoichiometry), which gave expression of the transfected gene 2-10-fold higher than the commercial reagent Lipofectin. At higher doses the new liposomes also caused markedly less cell death of KK-1 cells. On COS-7 cells these liposomes showed slightly, but significantly lower transfection, of approximately 70%, of that gained with Lipofectin. The murine Sertoli cells, MSC-1, selectively resisted transfection by the sphingosine derivative based liposomes tested, giving only 11-14% of the expression detected in Lipofectin transfected cells of the same line. In conclusion, the novel liposomes formulated offer an effective, technically easy and economical method of transfection for a variety of cultured cell lines.
Paukku
T
T
Department of Physiology, University of Turku, Finland. tommi.paukku@utu.fi
Lauraeus
S
S
Huhtaniemi
I
I
Kinnunen
P K
PK
D000818:Animals; D002412:Cations; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004075:Diglycerides / Q000032:analysis; D008081:Liposomes / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000032:analysis; D013110:Sphingosine / Q000031:analogs & derivatives / Q000032:analysis; D014162:Transfection
D002412:Cations; D004075:Diglycerides; D008081:Liposomes; D010714:Phosphatidylethanolamines; C044295:1,2-dioctanoylglycerol; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; C012491:phytosphingosine; D013110:Sphingosine