Fusion-mediated microinjection of liposome-enclosed DNA into cultured cells with the aid of influenza virus glycoproteins.
Influenza viruses were able to mediate fusion of DNA-loaded liposomes with living cultured cells such as monkey COS-7 cells. This was inferred from the appearance of CAT activity in recipient cells incubated with the combination of influenza viruses and liposomes loaded with the plasmid pSV2CAT. Influenza virions were found to be as efficient as intact Sendai virions in mediating microinjection of foreign DNA into living cells. Also, reconstituted envelopes bearing either influenza glycoproteins or the combination of Sendai and influenza glycoproteins were highly efficient in promoting fusion of loaded liposomes with recipient cells. Introduction of DNA into cultured cells required the presence of an active influenza fusion protein; namely, an active HA glycoprotein. Very little or no CAT activity was observed in cells incubated with loaded liposomes and unfusogenic influenza viruses. The virus-induced fusion event probably occurs within intracellular organelles such as endosomes following receptor-mediated endocytosis of virus-liposome complexes. This is due to the fact that the viral fusion glycoprotein is activated only at acidic pH values such as those which characterize the intraendosomal environment. Results of the present work demonstrate for the first time microinjection of foreign DNA via fusion with membranes of intracellular organelles. The potential of the present system to serve as a biological carrier for in vivo use is discussed.
Lapidot
M
M
Department of Biological Chemistry, Hebrew University of Jerusalem, Israel.
Loyter
A
A
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002784:Cholesterol; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004269:DNA, Bacterial / Q000008:administration & dosage / Q000235:genetics; D004337:Drug Carriers; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D009980:Influenza A virus; D008081:Liposomes; D008845:Microinjections / Q000379:methods; D010222:Parainfluenza Virus 1, Human; D010713:Phosphatidylcholines; D010957:Plasmids; D014759:Viral Envelope Proteins; D014771:Virion
D004269:DNA, Bacterial; D004337:Drug Carriers; D008081:Liposomes; D010713:Phosphatidylcholines; D014759:Viral Envelope Proteins; D004247:DNA; D002784:Cholesterol; D015500:Chloramphenicol O-Acetyltransferase