Synthesis and Enzymatic Characterization of Sugar-Modified Nucleoside Triphosphate Analogs

Stella Diafa (1) , Damien Evéquoz (1) , Christian Leumann (1) , Marcel Hollenstein (2)

1 UNIBE - Universität Bern / University of Bern
2 Chimie bioorganique des acides nucléiques - Bioorganic chemistry of nucleic acids

Chemical modification of nucleic acids can be achieved by the enzymatic polymerization of modified nucleoside triphosphates (dN*TPs). This approach obviates some of the requirements and drawbacks imposed by the more traditional solid-phase synthesis of oligonucleotides. Here, we describe the protocol that is necessary to synthesize dN*TPs and evaluate their substrate acceptance by polymerases for their subsequent use in various applications including selection experiments to identify aptamers. The protocol is exemplified for a sugar-constrained nucleoside analog, 7′,5′-bc-TTP.

Mots clés: Nucleoside triphosphate; Primer extension reactions; DNA polymerases; XNA; Solid-phase synthesis; Bicyclo-DNA

Domaines: Chimie; Chimie organique

Stella Diafa, Damien Evéquoz, Christian Leumann, Marcel Hollenstein. Synthesis and Enzymatic Characterization of Sugar-Modified Nucleoside Triphosphate Analogs. Non-Natural Nucleic Acids, 1973, Humana, pp.1-13, 2019, Methods in Molecular Biology, ⟨10.1007/978-1-4939-9216-4_1⟩. ⟨pasteur-03930944⟩