Gene transfer into hepatocytes using asialoglycoprotein receptor mediated endocytosis of DNA complexed with an artificial tetra-antennary galactose ligand

Bioconjug Chem. Nov-Dec 1992;3(6):533-9. doi: 10.1021/bc00018a012.


We have constructed an artificial ligand for the hepatocyte-specific asialoglycoprotein receptor for the purpose of generating a synthetic delivery system for DNA. This ligand has a tetra-antennary structure, containing four terminal galactose residues on a branched carrier peptide. The carbohydrate residues of this glycopeptide were introduced by reductive coupling of lactose to the alpha- and epsilon-amino groups of the two N-terminal lysines on the carrier peptide. The C-terminus of the peptide, containing a cysteine separated from the branched N-terminus by a 10 amino acid spacer sequence, was used for conjugation to 3-(2-pyridyldithio)propionate-modified polylysine via disulfide bond formation. Complexes containing plasmid DNA bound to these galactose-polylysine conjugates have been used for asialoglycoprotein receptor-mediated transfer of a luciferase gene into human (HepG2) and murine (BNL CL.2) hepatocyte cell lines. Gene transfer was strongly promoted when amphipathic peptides with pH-controlled membrane-disruption activity, derived from the N-terminal sequence of influenza virus hemagglutinin HA-2, were also present in these DNA complexes. Thus, we have essentially borrowed the small functional domains of two large proteins, asialoglycoprotein and hemagglutinin, and assembled them into a supramolecular complex to generate an efficient gene-transfer system.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Asialoglycoprotein Receptor
  • Asialoglycoproteins / metabolism
  • Cells, Cultured
  • DNA / chemistry
  • DNA / metabolism*
  • Endocytosis*
  • Galactose / chemistry
  • Galactose / metabolism*
  • Humans
  • Ligands
  • Liver / cytology
  • Liver / metabolism*
  • Luciferases / genetics
  • Mice
  • Molecular Sequence Data
  • Plasmids
  • Receptors, Immunologic / metabolism*
  • Transfection / methods*


  • Asialoglycoprotein Receptor
  • Asialoglycoproteins
  • Ligands
  • Receptors, Immunologic
  • DNA
  • Luciferases
  • Galactose