Immobilized Carbohydrates for Preparation of 3'-Glycoconjugated Oligonucleotides

Curr Protoc Nucleic Acid Chem. 2020 Dec;83(1):e122. doi: 10.1002/cpnc.122.

Abstract

A detailed protocol for preparation 3'-glycoconjugated oligonucleotides is described based on one-pot immobilization of 4,4'-dimethoxytrityl-protected carbohydrates to a solid support followed by on-support peracetylation and automated oligonucleotide assembly. Compared to an appropriate building block approach and post-synthetic manipulation of oligonucleotides, this protocol may simplify the synthesis scheme and increase overall yield of the conjugates. Furthermore, the immobilization to a solid support typically increases the stability of reactants, enabling prolonged storage, and makes subsequent processing convenient. Automated assembly on these carbohydrate-modified supports using conventional phosphoramidite chemistry produces 3'-glycoconjugated oligonucleotides in relatively high yield and purity. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Synthesis of 1-O-tert-butyldimethylsilyl-6-O-(4,4'-dimethoxytrityl)-β-D-glucose Basic Protocol 2: Synthesis of 6-O-dimethoxytrityl-2,3,1',3',4',6'-hexa-O-benzoylsucrose Basic Protocol 3: Synthesis of 6″-O-dimethoxytrityl-N-trifluoroacetyl-protected aminoglycosides Basic Protocol 4: Synthesis of 3-O-dimethoxytrityl-propyl β-D-galactopyranoside Basic Protocol 5: Synthesis of trivalent N-acetyl galactosamine cluster Basic Protocol 6: Synthesis of carbohydrate monosuccinates and their immobilization to a solid support Basic Protocol 7: Oligonucleotide synthesis using immobilized carbohydrates.

Keywords: carbohydrate oligonucleotide conjugates; delivery of oligonucleotides; solid-supported reactants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / chemistry
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Carbohydrates / chemistry*
  • Glycoconjugates / chemistry*
  • Oligonucleotides / chemistry*
  • Phosphoric Acids / chemistry
  • Spectrum Analysis

Substances

  • Amides
  • Carbohydrates
  • Glycoconjugates
  • Oligonucleotides
  • Phosphoric Acids
  • phosphoramidic acid