Sprecher
Beschreibung
The polymerization of modified nucleoside triphosphates (dNTPs) represents a versatile chemoenzymatic method for the introduction of chemical diversity into nucleic acids. These analogs have been employed in a variety of applications including functional tagging of oligonucleotides, formation of hydrogels, and de novo DNA synthesis. dNTPs also represent convenient vectors to expand the chemical diversity of oligonucleotide-based libraries in SELEX experiments for the generation of functional nucleic acids (i.e. catalysts and binders) with enhanced properties. In addition, dN*TPs equipped with suitable, transient 3’-protecting groups represent alluring synthons for controlled enzymatic synthesis of nucleic which in turn is used for the preparation of large amounts of oligonucleotides with view on storing digital information in DNA.
Here, I would like to summarize our recent efforts towards the synthesis of modified nucleoside triphosphates and their application in chemical biology. In a first section of this presentation, the recent identification of chemically modified aptamers will be presented (1, 2). A broad palette of chemical modifications can be used to promote binding activity and crystal structural analysis provide valuable information on the binding mechanism and the exact effect of these modifications on binding. The second part of this presentation will highlight our recent efforts towards the development of chemoenzymatic methods to produce modified oligonucleotides. This represents an unmet challenge in the field of aptamers and therapeutic oligonucleotides. We are currently exploring various approaches including the polymerization of transiently blocked nucleoside triphosphates by polymerases (3, 4) as well as ligation of short oligonucleotide fragments (5). We have also explored similar strategies to decorate oligonucleotides with triantennary GalNAc ligands which are key elements in therapeutic strategies to enhance specific delivery of nucleic acid drugs (6).
- Cheung, Y.-W.; Röthlisberger, P.; Mechaly, P. A. E.; Weber, P. W.; Levi-Acobas, F.; Lo, Y.; Wong, A. S. C.; Kinghorn, A. B.; Haouz, A.; Savage, G. P.; Hollenstein, M., Tanner, J. A., Evolution of abiotic cubane chemistries in a nucleic acid aptamer allows selective recognition of a malaria biomarker, Proc. Natl. Acad. Sci. U.S.A. 2020, 117, 16790-16798.
- Dahm, G. C.; Lim, L.; Akhtar, U.; Bouvier-Müller, A.; Levi-Acobas, F.; Bizat, P. N.; Niogret, G.; Tanner, J.; Ducongé, F.; Hollenstein, M.*, Probing antiaromatic cyclooctatetraene as nucleobase modification in aptamer selection, ChemRxiv 2025, doi:10.26434/chemrxiv-2025-cglxw.
- Pichon, M.; Levi-Acobas, F.; Kitoun, C.; Hollenstein, M.*, 2’,3’-protected nucleotides as building blocks for enzymatic de novo RNA synthesis, Chem. Eur. J. 2024, 30, e202400137.
- Sabat, N.; Katkevica, D.; Pajuste, K.; Flamme, M.; Stämpfli, A.; Katkevics, M.; Hanlon, S.; Marzuoli, I.; Bisagni, S.; Püntener, K.; Sladojevich, F.; Hollenstein, M.*, Towards the controlled enzymatic synthesis of LNA containing oligonucleotides, Front. Chem. 2023, 11, 1161462.
- Sabat, N.; Stämpfli, A.; Hanlon, S.; Marzuoli, I.; Bisagni, S.; Sladojevich, F.; Püntener, K.; Hollenstein, M.*, Template-dependent DNA ligation for the synthesis of modified oligonucleotides, Nat. Commun. 2024, 15, 8009.
- Dhara, D.; Mulard, L.; Hollenstein, M.*, Role of Carbohydrate in Nucleic Acids, Chem. Soc. Rev. 2025, 54, 2948–2983.
