NBD-conjugated biosurfactant (MEL-A) shows a new pathway for transfection.
Gene transfection is a fundamental technology for molecular and cell biology, and also clinical gene therapy. A variety of non-viral vectors have been investigated for gene transfection, but their gene delivery had remained an inefficient process. Recently, we found that a biosurfactant, mannosylerythritol lipid (MEL)-A, dramatically increased the efficiency in transfection of plasmid DNA mediated by cationic liposomes. However, its mechanism has not been understood yet. Here we examined the mechanism of the transfection mediated by cationic liposomes with NBD-conjugated MEL-A. We found that MEL-A first gradually distributed on the intracellular membranes through the plasma membranes of target cells, while the cationic liposomes with MEL-A fused to the plasma membranes in 20-35 min. Thereafter, the oligonucleotide released from the vesicles was immediately transferred to the nucleus. The present results showed a new role of non-viral vectors in transfection.
Ueno
Yoshinobu
Y
Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan.
Inoh
Yoshikazu
Y
Furuno
Tadahide
T
Hirashima
Naohide
N
Kitamoto
Dai
D
Nakanishi
Mamoru
M
D021581:Active Transport, Cell Nucleus; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D002412:Cations; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004705:Endocytosis; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D017930:Genes, Reporter; D006017:Glycolipids / Q000737:chemistry*; D007425:Intracellular Membranes / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D008081:Liposomes; D008156:Luciferases / Q000235:genetics; D008561:Membrane Fusion; D051379:Mice; D008958:Models, Molecular; D008968:Molecular Conformation; D041681:NIH 3T3 Cells; D009578:Nitrobenzenes / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism*; D010069:Oxadiazoles / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D000077191:Wortmannin
D000730:Androstadienes; D002412:Cations; D005456:Fluorescent Dyes; D006017:Glycolipids; D008055:Lipids; D008081:Liposomes; D009578:Nitrobenzenes; D009841:Oligonucleotides; D010069:Oxadiazoles; D013501:Surface-Active Agents; C439671:mannosylerythritol lipid; D004247:DNA; D008156:Luciferases; D016650:Fluorescein-5-isothiocyanate; D000077191:Wortmannin