Protamine-modified DDAB lipid vesicles promote gene transfer in the presence of serum.
Cationic lipid vesicle-mediated gene transfer has become common for in vitro gene delivery. However, the transfection efficiency is often impaired by serum. DDAB (dimethyldioctadecyl ammonium bromide) lipid vesicle-mediated gene transfer, which we previously reported, has the same problem. To overcome this obstacle, we here report a novel transfection vehicle using protamine-modified DDAB lipid vesicles. While free protamine was simply added to the DNA/lipid complex in the previous study, in the present method the protamine is chemically conjugated to stearic acid and incorporated into DDAB lipid vesicles. Gene transfer was not significantly inhibited in 10% serum-containing medium by this method for the transfection of cultured cells. Protamine-modified DDAB lipid vesicles also enhanced virus transduction efficiency in the presence of serum using a replication-defective retroviral vector. Furthermore, the vesicles allowed efficient gene transfer for avian embryos in vivo. These results indicate that the method is useful for the production of transgenic animals and gene therapy.
Mizuarai
S
S
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Chikusa-ku, Nagoya 464-8603, Japan.
Ono
K
K
You
J
J
Kamihira
M
M
Iijima
S
S
D016475:3T3 Cells; D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D006224:Cricetinae; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004622:Embryo, Mammalian / Q000502:physiology; D004625:Embryo, Nonmammalian; D018014:Gene Transfer Techniques; D051379:Mice; D011479:Protamines / Q000737:chemistry*; D011784:Quail; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D012190:Retroviridae / Q000235:genetics; D014162:Transfection / Q000379:methods
D011479:Protamines; D000644:Quaternary Ammonium Compounds; C046112:didodecyldimethylammonium; D004247:DNA