Get access

Efficient Solid-Phase Synthesis of pppRNA by Using Product-Specific Labeling

Authors

  • M. Goldeck,

    1. Institute for Clinical Chemistry & Clinical Pharmacology, University Hospital Bonn, Sigmund-Freud-Strasse 25, 53127 Bonn (Germany)
    Search for more papers by this author
  • Prof. Dr. T. Tuschl,

    1. Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, Box 186, New York, NY 10065 (USA)
    Search for more papers by this author
  • Prof. Dr. G. Hartmann,

    1. Institute for Clinical Chemistry & Clinical Pharmacology, University Hospital Bonn, Sigmund-Freud-Strasse 25, 53127 Bonn (Germany)
    Search for more papers by this author
  • Dr. J. Ludwig

    Corresponding author
    1. Institute for Clinical Chemistry & Clinical Pharmacology, University Hospital Bonn, Sigmund-Freud-Strasse 25, 53127 Bonn (Germany)
    • Institute for Clinical Chemistry & Clinical Pharmacology, University Hospital Bonn, Sigmund-Freud-Strasse 25, 53127 Bonn (Germany)===

    Search for more papers by this author

  • This research was supported by grants from the German Research Foundation (SFB670, SFB704) to G.H. M.G., G.H., and J.L. are funded by the excellence cluster Immunosensation. We thank Dr. I. Roehl and S. Seiffert (Axolabs GmbH, Analytics) for measurement of the LC–MS spectra and helpful discussions. pppRNA=5′-triphosphate RNA.

Abstract

A novel solid-phase synthesis and purification strategy for 5′-triphosphate oligonucleotides by using lipophilic tagging of the triphosphate moiety is reported. This is based on triphosphate synthesis with 5′-O-cyclotriphosphate intermediates, whereby a lipophilic tag, such as decylamine, is introduced during the ring-opening reaction to give a linear gamma-phosphate-tagged species. This method enables the highly efficient synthesis of 5′-triphosphorylated RNA derivatives and their gamma-phosphate-substituted analogues and will especially facilitate the advancement of therapeutic approaches that make use of 5′-triphosphate oligonucleotides as potent activators of the cytosolic immune sensor RIG-I.

Ancillary