Construction of a multifunctional envelope-type nano device by a SUV*-fusion method.

A novel assembly method "SUV*-fusion method" was developed for the construction of a small and homogenous multifunctional envelope-type nano device (MEND) by utilizing a detergent-rich small unilamellar vesicle (SUV*). The method consists of three steps: (1) DNA condensation with a polycation, (2) electrostatic interaction of the SUV* with the DNA/polycation complex (DPC) and (3) lipid coating of DPC by SUV* fusion via removal of the detergent. We confirmed the construction of the MEND by sucrose density gradient centrifugation, and isolated the MEND only from the boundary between 25% and 40% sucrose. The isolated MEND had a small diameter (155 nm), was negatively charged (-24 mV), and encapsulated 30% of the total DNA. The MEND was formed by only SUV*, not by a lipid/detergent micelle. This confirms that a small and homogenous MEND can be constructed by the SUV*-fusion method. Furthermore, we confirmed that a transferrin-modified MEND could deliver a gene into a cell through receptor-mediated endocytosis. Consequently, we report on the successful construction of a small and homogenous MEND by a novel SUV*-fusion method.

[1]  I. Maclachlan,et al.  Stabilized plasmid-lipid particles: a systemic gene therapy vector. , 2002, Methods in enzymology.

[2]  Shiroh Futaki,et al.  Development of a non-viral multifunctional envelope-type nano device by a novel lipid film hydration method. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[3]  H. Harashima,et al.  Visualization of intracellular trafficking of exogenous DNA delivered by cationic liposomes. , 2002, Biochemical and biophysical research communications.

[4]  Kazuo Maruyama,et al.  Transferrin-modified liposomes equipped with a pH-sensitive fusogenic peptide: an artificial viral-like delivery system. , 2004, Biochemistry.

[5]  H Harashima,et al.  Mechanism of improved gene transfer by the N-terminal stearylation of octaarginine: enhanced cellular association by hydrophobic core formation , 2004, Gene Therapy.

[6]  N. Minakawa,et al.  No enhancement of nuclear entry by direct conjugation of a nuclear localization signal peptide to linearized DNA. , 2003, Bioconjugate chemistry.

[7]  Y. Shinohara,et al.  Intracellular control of gene trafficking using liposomes as drug carriers. , 2001, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[8]  W. Pardridge,et al.  Brain-specific expression of an exogenous gene after i.v. administration , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[9]  R D Klausner,et al.  Receptor-mediated endocytosis of transferrin in K562 cells. , 1983, The Journal of biological chemistry.

[10]  Hideyoshi Harashima,et al.  Pharmacokinetic and pharmacodynamic considerations in gene therapy. , 2003, Drug discovery today.

[11]  L. Palmer,et al.  Stabilized plasmid-lipid particles: construction and characterization , 1999, Gene Therapy.

[12]  H Akita,et al.  Quantitative three-dimensional analysis of the intracellular trafficking of plasmid DNA transfected by a nonviral gene delivery system using confocal laser scanning microscopy. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[13]  G Gregoriadis,et al.  Vaccine entrapment in liposomes. , 1999, Methods.

[14]  M. Ueno.  Partition behavior of a nonionic detergent, octyl glucoside, between membrane and water phases, and its effect on membrane permeability. , 1989, Biochemistry.

[15]  G. Gregoriadis,et al.  Liposome-entrapped plasmid DNA: characterisation studies. , 2000, Biochimica et biophysica acta.

[16]  T. Suhara,et al.  Factors governing the in vivo tissue uptake of transferrin-coupled polyethylene glycol liposomes in vivo. , 2004, International journal of pharmaceutics.