Immobilized Carbohydrates for Preparation of 3′‐Glycoconjugated Oligonucleotides

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

Controlled Monofunctionalization of Molecular Spherical Nucleic Acids on a Buckminster Fullerene Core

“…3-Phenyl 1,2,4-dithiazoline-5-one (POS) was used as a sulfurization reagent for the synthesis of ON5 . Our previously reported customized solid supports , were utilized for the synthesis of appropriately 3′-modified ONs: ON7 and ON10 with a 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) and a d -galactose moiety, respectively. While ON1 and ON2 are short sequences, preliminarily used to demonstrate the SNA’s assembly ( S1 and S2 ), ON3 – ON10 are biologically active sequences.…”