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Nucleic Acids Symposium Series 2001 1(1):217-218; doi:10.1093/nass/1.1.217
© 2001 by Oxford University Press
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Protein/oligonucleotide conjugates as a cell specific PNA carrier

Kentaro Obara, Tsutomu Ishihara, Toshihiro Akaike and Atsushi Maruyama

Department of Biomolecular Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori,Yokohama 226-8501, Japan

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 reseptor (ASGP-R) mediated endocytosis to be a major-route for the cellular uptake.


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