Evaluation of 3′-phosphate as a transient protecting group for controlled enzymatic synthesis of DNA and XNA oligonucleotides

Autor: Marie Flamme, Steven Hanlon, Irene Marzuoli, Kurt Püntener, Filippo Sladojevich, Marcel Hollenstein
Přispěvatelé: Institut Pasteur [Paris] (IP), Chimie bioorganique des acides nucléiques - Bioorganic chemistry of nucleic acids, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), F. Hoffmann-La Roche [Basel], The authors thank Institut Pasteur (starting funds to M.H.) and Roche for financial support. We would like to thank Siegfried Stolz and Andreas Stämpfli (Roche) for help with the LCMS analysis and Martin Olbrich (Roche) for help with the manuscript preparation and for fruitful discussions. We would like to thank Chiara Figazzolo (Institut Pasteur) for running additional HPLC experiments, Olena Mayboroda and Sébastien Brier (Institut Pasteur) for the MS analysis of the Klenow polymerase.
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Communications Chemistry
Communications Chemistry, 2022, 5 (1), pp.68. ⟨10.1038/s42004-022-00685-5⟩
ISSN: 2399-3669
Popis: Chemically modified oligonucleotides have advanced as important therapeutic tools as reflected by the recent advent of mRNA vaccines and the FDA-approval of various siRNA and antisense oligonucleotides. These sequences are typically accessed by solid-phase synthesis which despite numerous advantages is restricted to short sequences and displays a limited tolerance to functional groups. Controlled enzymatic synthesis is an emerging alternative synthetic methodology that circumvents the limitations of traditional solid-phase synthesis. So far, most approaches strived to improve controlled enzymatic synthesis of canonical DNA and no potential routes to access xenonucleic acids (XNAs) have been reported. In this context, we have investigated the possibility of using phosphate as a transient protecting group for controlled enzymatic synthesis of DNA and locked nucleic acid (LNA) oligonucleotides. Phosphate is ubiquitously employed in natural systems and we demonstrate that this group displays most characteristics required for controlled enzymatic synthesis. We have devised robust synthetic pathways leading to these challenging compounds and we have discovered a hitherto unknown phosphatase activity of various DNA polymerases. These findings open up directions for the design of protected DNA and XNA nucleoside triphosphates for controlled enzymatic synthesis of chemically modified nucleic acids.
Databáze: OpenAIRE
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