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J. Biochem, 2001, Vol. 129, No. 1 125-132
© 2001 Japanese Biochemical Society

other

Protamine-Modified DDAB Lipid Vesicles Promote Gene Transfer in the Presence of Serum1

Shinji Mizuarai, Ken-ichiro Ono, Jun You, Masamichi Kamihira2 and Shinji Iijima

Department of Biotechnology, Graduate School of Engineering, Nagoya University Chikusa-ku, Nagoya 464-8603

2 To whom correspondence should be addressed. Tel: +81-52-789-4277, Fax: +81-52-789-3221, E-mail: kamihira{at}proc.nubio.nagoya-u.ac.jp

Cationic lipid vesicle-mediated gene transfer has become common for in vitro gene delivery. However, the transfection efficiency is often impaired by serum. DDAB (dime-thyldioctadecyl 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.

1This work was supported in part by Grants-in-Aid for Scientific Research from Ministry of Education, Science, Sports and Culture of Japan (Nos. 10145104 and 12555227).


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[Abstract] [PDF]


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