Synthesis, cell-surface binding, and cellular uptake of fluorescently labeled glucose-DNA conjugates with different carbohydrate presentation

Bioconjug Chem. 2010 Jul 21;21(7):1280-7. doi: 10.1021/bc100079n.

Abstract

Oligonucleotide conjugates carrying carbohydrates at the 5'-end have been prepared. Glucose, fucose, and saccharides containing glucose at the nonreducing end were attached to DNA strands using the classical phosphoramidite chemistry. Two types of spacers and a dendron scaffold helped to obtain a diversity of sugar presentations in the DNA conjugates. Cellular surface adsorption and cellular uptake of carbohydrate oligonucleotide antisense sequences were measured using flow cytometric analysis. Conjugates with the glucose moiety linked through long spacers (15 to 18 atom distances) were internalized better than those with short linkers (4 atom distance) and than DNA control strands without sugar modification. Conjugates with tetravalent presentation of glucose did not improve cell uptake.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Binding Sites
  • Biological Transport
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • DNA / chemistry
  • DNA / metabolism*
  • Flow Cytometry
  • Fluorescent Dyes / chemistry*
  • Glucose / chemistry
  • Glucose / metabolism*
  • Glycoconjugates / chemical synthesis*
  • Glycoconjugates / chemistry
  • Glycoconjugates / metabolism*
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence
  • Molecular Structure
  • Oligonucleotides / chemistry
  • Organophosphorus Compounds / chemistry
  • Stereoisomerism

Substances

  • Fluorescent Dyes
  • Glycoconjugates
  • Oligonucleotides
  • Organophosphorus Compounds
  • phosphoramidite
  • DNA
  • Glucose