Synthesis of Dinucleotide Non‐Symmetrical Triester Phosphate Phosphoramidites and Their Incorporation Within Oligonucleotides

Abstract

AbstractIn this protocol article, the synthesis of dinucleotide non‐symmetrical triester phosphate phosphoramidites will be highlighted. Specifically, we use a selective transesterification starting with tris(2,2,2‐trifluoroethyl) phosphate to afford a dinucleotide derivative phosphate ester. Substitution of the final trifluoroethyl group with various alcohols affords a dinucleotide triester phosphate with a hydrophobic group, which can then be deprotected and converted to a phosphoramidite for incorporation within oligonucleotides. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Synthesis of a DMT‐ and TBS‐protected unsymmetrical dinucleotideBasic Protocol 2: Synthesis of a DMT‐protected unsymmetrical dinucleotide phosphotriester monoalcoholsBasic Protocol 3: Synthesis of DMT‐protected phenylethyl phosphotriester dinucleotide phosphoramiditesBasic Protocol 4: Synthesis, purification, and characterization of RNAs containing triester phosphate modifications