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-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 10(3) 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 10(1)-10(2) 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.
Schuster
M J
MJ
Department of Medicine, Division of Gastroenterology-Hepatology, University of Connecticut School of Medicine, Farmington, Connecticut 06030, USA.
Wu
G Y
GY
Walton
C M
CM
Wu
C H
CH
D000818:Animals; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins / Q000737:chemistry*; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000008:administration & dosage* / Q000378:metabolism; D004337:Drug Carriers; D004705:Endocytosis; D005260:Female; D019204:GTP-Binding Proteins / Q000737:chemistry*; D015870:Gene Expression; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D009961:Orosomucoid / Q000031:analogs & derivatives* / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000737:chemistry*
D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D004337:Drug Carriers; D009961:Orosomucoid; D011956:Receptors, Cell Surface; C017030:asialoorosomucoid; D011107:Polylysine; D004247:DNA; D019204:GTP-Binding Proteins