Synthesis of stereospecific oligonucleotide phosphorothioates

C - Chemistry – Metallurgy – 12 – P

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C12P 19/34 (2006.01) C07H 21/04 (2006.01) C12N 15/11 (2006.01)

Patent

CA 2208528

Disclosed is a method for synthesizing of stereospecific (Rp) phosphorothioate oligonucleotides. In this method, a primer comprising a plurality of deoxyribonucleotides and a ribonucleotide at the 5' terminal or 5' penultimate position, is annealed to a template. The structure is contacted with a mixture of deoxynucleoside .alpha.-triphosphate Sp diastereomers and a DNA polymerase to form a PS-Rp oligodeoxynucleotide extension which is liberated as a single- stranded PS-Rp oligonucleotide by cleavage after the ribonucleotide in the primer. Also disclosed are PS-Rp oligonucleotides and oligonucleotides prepared according to this method.

L'invention concerne un procédé de synthèse d'oligonucléotides de phosphorothioates stéréospécifiques (Rp). Ce procédé consiste à hybrider à une matrice une amorce composée d'une pluralité de désoxyribonucléotides et d'un ribonucléotide au niveau de la position terminale 5' ou de la position pénultième 5'. Cette structure est mise en contact avec un mélange de diastéréomères Sp de .alpha.-triphosphate de désoxynucléosides et d'une ADN polymérase afin qu'une extension d'oligodeoxynucléotides PS-Rp soit formée et libérée en tant qu'oligonucléotide monocaténaire PS-Rp par coupure après le ribonucléotide de l'amorce. L'invention concerne également des oligonucléotides PS-Rp, ainsi que des oligonucléotides préparés selon ce procédé.

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