A Versatile and Convenient Synthesis of 34 S-Labeled Phosphorothioate Oligonucleotides

Chembiochem. 2018 Oct 4;19(19):2114-2119. doi: 10.1002/cbic.201800417. Epub 2018 Sep 13.

Abstract

A synthetic protocol for 34 S-labeled phosphorothioate oligonucleotides (PS ONs) was developed to facilitate MS-based assay analysis. This was enabled by a highly efficient, two-step, one-pot synthesis of 34 S-labeled phenylacetyl disulfide (34 S-PADS), starting from 34 S-enriched elemental sulfur (34 S8 ). 34 S-PADS was subsequently used for stable isotope labeling (SIL) of oligonucleotides containing a phosphorothioate backbone. The 34 S-SIL PS ONs are shown to retain the same melting temperature, antisense activity, and secondary structure as those of the corresponding unlabeled 32 S PS ONs.

Keywords: antisense agents; isotopic labeling; oligonucleotides; sulfur; synthesis design.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Glucagon-Like Peptide-1 Receptor / metabolism
  • HEK293 Cells
  • Humans
  • Isotope Labeling
  • Oligonucleotides, Antisense* / chemical synthesis
  • Oligonucleotides, Antisense* / chemistry
  • Phenylacetates* / chemical synthesis
  • Phenylacetates* / chemistry
  • Phosphorothioate Oligonucleotides* / chemical synthesis
  • Phosphorothioate Oligonucleotides* / chemistry
  • RNA, Long Noncoding / metabolism
  • Sulfides* / chemical synthesis
  • Sulfides* / chemistry
  • Sulfur / chemistry

Substances

  • GLP1R protein, human
  • Glucagon-Like Peptide-1 Receptor
  • MALAT1 long non-coding RNA, human
  • Oligonucleotides, Antisense
  • Phenylacetates
  • Phosphorothioate Oligonucleotides
  • RNA, Long Noncoding
  • Sulfides
  • phenylacetyl disulfide
  • Sulfur