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Rapid Synthesis of Nucleoside Triphosphates and Analogues

Ripp, A; Singh, J; Jessen, HJ; (2020) Rapid Synthesis of Nucleoside Triphosphates and Analogues. Current Protocols in Nucleic Acid Chemistry , 81 (1) , Article e108. 10.1002/cpnc.108. Green open access

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Abstract

Nucleoside triphosphates (NTPs) are essential biomolecules involved in almost all biological processes, and their study is therefore critical to understanding cellular biology. Here, we describe a chemical synthesis suitable for obtaining both natural and highly modified NTPs, which can, for example, be used as surrogates to probe biological processes. The approach includes the preparation of a reagent that enables the facile introduction and modification of three phosphate units: cyclic pyrophosphoryl P‐amidite (c‐PyPA), derived from pyrophosphate (PV) and a reactive phosphoramidite (PIII). By using non‐hydrolyzable analogues of pyrophosphate, the reagent can be readily modified to obtain a family of non‐hydrolyzable analogues containing CH2, CF2, CCl2, and NH that are stable in solution for several weeks if stored appropriately. They enable the synthesis of NTPs by reaction with nucleosides to give deoxycyclotriphosphate esters that are then oxidized to cyclotriphosphate (cyclo‐TP) esters. The use of different oxidizing agents provides an opportunity for modification at P‐α. Furthermore, terminal modifications at P‐γ can be introduced by linearization of the cyclo‐TP ester with various nucleophiles. / Basic Protocol 1: Synthesis of cyclic pyrophosphoryl P‐amidite (c‐PyPA) and derivatives (c‐PyNHPA, c‐PyCH2PA, c‐PyCCl2PA, c‐PyCF2PA) / Basic Protocol 2: Synthesis of 3′‐azidothymidine 5′‐γ‐P‐propargylamido triphosphates and analogues / Basic Protocol 3: Synthesis of 2′‐deoxythymidine 5′‐γ‐P‐propargylamido triphosphate (15) / Basic Protocol 4: Synthesis of adenosine 5′‐γ‐P‐amido triphosphate (19) and adenosine 5′‐γ‐P‐propargylamido triphosphate (20) / Basic Protocol 5: Synthesis of d4T 5′‐γ‐propargylamido β,γ‐(difluoromethylene)triphosphate / Support Protocol: Synthesis of diisopropylphosphoramidous dichloride

Type: Article
Title: Rapid Synthesis of Nucleoside Triphosphates and Analogues
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/cpnc.108
Publisher version: https://doi.org/10.1002/cpnc.108
Language: English
Additional information: Copyright © 2020 The Authors. This is an open access article under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: cyclotriphosphate, non-hydrolyzable nucleotide analogues, nucleoside 5′-triphosphate, phosphoramidite, phosphorylation
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > Dept of Chemistry
URI: https://discovery.ucl.ac.uk/id/eprint/10113395
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