Gene delivery into hepatocytes with the preS/liposome/DNA system.
Gene delivery into human hepatocytes remains a critical issue for the development of liver-directed gene therapy. Gene delivery based on non-viral vectors is an attractive approach relative to viral vectors. In this report, novel delivery system of preS/liposome/DNA virus-like particle (VLP) was developed for gene transfection into hepatocytes in vivo and in vitro. Plasmid pCMVbeta, expressing beta-galactosidase, was encapsulated with cationic liposome, and then the histidine-tagged preS domain of hepatitis B virus was coated on the surface of liposome/DNA to form preS/liposome/ DNA VLP. Transfection efficiencies of preS/liposome/DNA, liposome/DNA, naked DNA and preS were analyzed using several different human cell lines. The highest transfection efficiency was found using preS/liposome/DNA VLP as the transfection reagent in human hepatocyte (HH) cell line. Results show that preS domain of hepatitis B virus coated on liposome/DNA can be used for highly efficient gene transfection into human hepatocytes. Moreover, the target characteristic of preS/liposome/DNA was analyzed in vivo. After preS/liposome/DNA VLP was injected into immunocompromised (Nude) mice via the tail vein, most of beta-galactosidase was expressed in the liver; however, no significant target expression was found with the injection of liposome/ DNA or naked DNA. Our results show that preS/liposome/DNA VLP can be used as a novel liver-specific gene delivery system.
Wang
Zhijun
Z
CAS-MPG Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, People's Republic of China. zjwang@picb.ac.cn
Yuan
Zhenghong
Z
Jin
Li
L
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007275:Injections, Intravenous; D008081:Liposomes / Q000737:chemistry*; D051379:Mice; D008819:Mice, Nude; D010957:Plasmids / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
D008081:Liposomes; D011498:Protein Precursors; D014759:Viral Envelope Proteins; C101581:preS (120-145) peptide; D004247:DNA