Effect of lipid compositions on gene transfer into 293 cells using Sendai F/HN-virosomes.
Fusogenic liposomes that incorporate Sendai virus envelope proteins, so-called Sendai virosomes, have been developed for in vitro and in vivo genetic modification of animal cells. In this study, several different virosomes of varying lipid compositions were formulated and their in vitro gene-transfer efficiencies compared. The virosomes were prepared by quantitative reconstitution of the Sendai envelope, fusion (F) and hemagglutinin-neuraminidase (HN) proteins into liposomal vesicles. Virosomes that contained luciferase reporter genes were tested in 293 transformed human kidney cells. F/HN-virosomes that were prepared with an artificial Sendai viral envelope (ASVE-virosomes) or phosphatidylserine (PS-virosomes) exhibited an 8- or 6-fold higher gene-transfer efficiency than cationic liposomes that were made with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). F/HNvirosomes that were prepared with phosphatidic acid (PA-virosomes) instead of PS were less efficient in gene transfer than either ASVE- or PS-virosomes. In addition, the gene-transfer capability of ASVE- and PS-virosomes was maximal at a Ca2+ concentration of 510 mM. These results suggest that the incorporated lipid components significantly affect the in vitro gene transfer that is mediated by Sendai F/HN-virosomes.
D002118:Calcium / Q000378:metabolism; D002461:Cell Line, Transformed; D005822:Genetic Vectors / Q000378:metabolism / Q000648:ultrastructure; D005960:Glucosides / Q000378:metabolism; D015707:HN Protein / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism; D008156:Luciferases / Q000378:metabolism; D029082:Sendai virus / Q000378:metabolism*; D014162:Transfection / Q000379:methods*; D014760:Viral Fusion Proteins / Q000378:metabolism; D014763:Viral Matrix Proteins / Q000378:metabolism; D022701:Virosomes / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure
D005960:Glucosides; D015707:HN Protein; D014760:Viral Fusion Proteins; D014763:Viral Matrix Proteins; D022701:Virosomes; C018619:octyl-beta-D-glucoside; D008156:Luciferases; D002118:Calcium