Protein/oligonucleotide conjugates as a cell specific PNA carrier.
We have focused on proteineus ligand conjugate with oligonucleotides (ODNs) as a cell-specific delivery vector for peptide nucleic acids (PNAs). Asialofetuin (AF), a hepatocyte-specific proteineus ligand, was conjugated with ODNs that served as binding sites for PNAs. Succinimidyl-transe-4(N-maleimidylmethyl)-cyclohexane-1-carboxylate (SMCC) modified AF was coupled with 5'-thiolated oligodeoxynucleotide (HS-ODN). The resulting conjugate held PNAs with sequence-specific manner. The PNA/DNA conjugate complex has resistance against nucleases in serum. The efficient release of PNA from the complex was observed when the complex was made in contact with a target nucleotide. PNA uptake to hepatocytes was greatly enhanced when hepatocytes was incubated with PNA/conjugate complex. Free AF thoroughly inhibited PNA uptake with the conjugate, evidencing asialoglycoprotein receptor (ASGP-R) mediated endocytosis to be a major-route for the cellular uptake.
Obara
K
K
Department of Biomolecular Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori, Yokohama 226-8501, Japan.
Ishihara
T
T
Akaike
T
T
Maruyama
A
A
D000818:Animals; D001212:Asialoglycoproteins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002478:Cells, Cultured; D004337:Drug Carriers; D004705:Endocytosis; D060748:Fetuins; D022781:Hepatocytes / Q000378:metabolism; D051379:Mice; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism*; D020135:Peptide Nucleic Acids / Q000378:metabolism*; D000509:alpha-Fetoproteins / Q000737:chemistry / Q000378:metabolism*
D001212:Asialoglycoproteins; D004337:Drug Carriers; D060748:Fetuins; D009838:Oligodeoxyribonucleotides; D020135:Peptide Nucleic Acids; D000509:alpha-Fetoproteins; C017029:asialofetuin