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Updated: May 30, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Synthesis of oligoribonucleotides with phosphonate-modified linkages
Ondřej Páv1, Ivana Košiová, Ivan Barvík
1Institute of Organic Chemistry and Biochemistry AS CR, v.v.i., Flemingovo nam. 2, 16610, Prague, Czech Republic.
Abstract:
Solid phase synthesis of phosphonate-modified oligoribonucleotides using 2'-O-benzoyloxymethoxymethyl protected monomers is presented in both 3'→5' and 5'→3' directions. Hybridisation properties and enzymatic stability of oligoribonucleotides modified by regioisomeric 3'- and 5'-phosphonate linkages are evaluated. The introduction of the 5'-phosphonate units resulted in moderate destabilisation of the RNA/RNA duplexes (ΔT(m)-1.8 °C/mod.), whereas the introduction of the 3'-phosphonate units resulted in considerable destabilisation of the duplexes (ΔT(m)-5.7 °C/mod.). Molecular dynamics simulations have been used to explain this behaviour. Both types of phosphonate linkages exhibited remarkable resistance in the presence of ribonuclease A, phosphodiesterase I and phosphodiesterase II.
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