Multicomponent DNA Carrier with a Vesicular Stomatitis Virus G-Peptide Greatly Enhances Liver-Targeted Gene Expression in Mice

Genes can be targeted to hepatocytes in vitro and in vivo by the use of asialoorosomucoid−polylysine conjugates. After systemic application, this nonviral vector is recognized by highly selective asialoglycoprotein (AsGP) receptors on the sinusoidal liver cell membrane and is taken up via receptor-m...

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Bibliographic Details
Published in:Bioconjugate chemistry 1999-11, Vol.10 (6), p.1075-1083
Main Authors: Schuster, Martin J, Wu, George Y, Walton, Cherie M, Wu, Catherine H
Format: Article
Language:English
Subjects:
Animals
Asialoglycoprotein Receptor
Asialoglycoproteins - chemistry
Cell Membrane - metabolism
Cell Nucleus - metabolism
DNA - administration & dosage
DNA - metabolism
Drug Carriers
Endocytosis
Female
Gene Expression
Gene Targeting
Gene Transfer Techniques
GTP-Binding Proteins - chemistry
Liver - drug effects
Liver - metabolism
Liver - ultrastructure
Mice
Mice, Inbred BALB C
Orosomucoid - analogs & derivatives
Orosomucoid - chemistry
Polylysine - chemistry
Receptors, Cell Surface - metabolism
Vesicular stomatitis Indiana virus - chemistry
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Summary:Genes can be targeted to hepatocytes in vitro and in vivo by the use of asialoorosomucoid−polylysine conjugates. After systemic application, this nonviral vector is recognized by highly selective asialoglycoprotein (AsGP) receptors on the sinusoidal liver cell membrane and is taken up via receptor-mediated endocytosis. As most of the DNA is rapidly transferred to lysosomes where it is degraded, transfection efficiency is low and gene expression transient. To address this problem, we incorporated a pH-dependent synthetic hemolytic peptide derived of the G-protein of Vesicular Stomatitis Virus (VSV) into the gene transfer system, to increase endosomal escape of internalized DNA. The multicomponent carrier binds DNA in a nondamaging way, is still recognized by the AsGP receptor, and is targeted to the liver in vivo. Injection of DNA complexes containing a luciferase marker gene resulted in luciferase expression of 29 000 pg/g liver which corresponded to an increase of a factor of 103 overexpression after injection of DNA complexes without endosomolytic peptide. Furthermore, the amount of intact transgene within isolated liver cell nuclei was increased by a factor of 101−102 by the use of the multicomponent carriers. These results demonstrate that incorporation of a hemolytic peptide into a nonviral vector can greatly increase gene expression while retaining cell type targetability in vivo.
ISSN:1043-1802
1520-4812
DOI:10.1021/bc990071r