Enzymatic Synthesis of Glyco-DNA Equipped with Oligosaccharides and Charged Monosaccharides

) bearing mono- (either neutral or charged moieties) and oligosaccharides affixed to position 5 of the nucleobase. The resulting glyco-modified nucleotides were well incorporated into DNA by different template-dependent polymerases, but much less by template-independent polymerases. While the presence of charged residues did not affect incorporation efficiency, glycan side-chain extension considerably reduced the incorporation yield of the glycosylated nucleotides. On the other hand, the saccharide appendages did not affect the serum stability of the 71-mer sequences containing the glyco-modified dUTPs and were only slightly destabilizing duplex DNA. Interestingly, the presence of short oligosaccharides highly increased the oligonucleotide stability against the BfaI endonuclease. These findings show that 5-glycosylation with short oligosaccharides could be a strategy to expand the chemical arsenal on nucleobases of DNA, offering a robust strategy for developing next-generation glyco-aptamers with enhanced endonuclease stability and novel saccharide-mediated recognition capabilities.