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In Vivo Incorporation of Azide Groups into DNA by Using Membrane-Permeable Nucleotide Triesters


Tera, Masayuki; Glasauer, Stella M K; Luedtke, Nathan W (2018). In Vivo Incorporation of Azide Groups into DNA by Using Membrane-Permeable Nucleotide Triesters. Chembiochem, 19(18):1939-1943.

Abstract

Metabolic incorporation of bioorthogonal functional groups into cellular nucleic acids can be impeded by insufficient phosphorylation of nucleosides. Previous studies found that 5azidomethyl‐2′‐deoxyuridine (AmdU) was incorporated into the DNA of HeLa cells expressing a low‐fidelity thymidine kinase, but not by wild‐type HeLa cells. Here we report that membrane‐permeable phosphotriester derivatives of AmdU can exhibit enhanced incorporation into the DNA of wild‐type cells and animals. AmdU monophosphate derivatives bearing either 5′‐bispivaloyloxymethyl (POM), 5′‐bis‐(4‐acetoxybenzyl) (AB), or “Protide” protective groups were used to mask the phosphate group of AmdU prior to its entry into cells. The POM derivative “POM‐AmdU” exhibited better chemical stability, greater metabolic incorporation efficiency, and lower toxicity than “AB‐AmdU”. Remarkably, the addition of POM‐AmdU to the water of zebrafish larvae enabled the biosynthesis of azide‐modified DNA throughout the body.

Abstract

Metabolic incorporation of bioorthogonal functional groups into cellular nucleic acids can be impeded by insufficient phosphorylation of nucleosides. Previous studies found that 5azidomethyl‐2′‐deoxyuridine (AmdU) was incorporated into the DNA of HeLa cells expressing a low‐fidelity thymidine kinase, but not by wild‐type HeLa cells. Here we report that membrane‐permeable phosphotriester derivatives of AmdU can exhibit enhanced incorporation into the DNA of wild‐type cells and animals. AmdU monophosphate derivatives bearing either 5′‐bispivaloyloxymethyl (POM), 5′‐bis‐(4‐acetoxybenzyl) (AB), or “Protide” protective groups were used to mask the phosphate group of AmdU prior to its entry into cells. The POM derivative “POM‐AmdU” exhibited better chemical stability, greater metabolic incorporation efficiency, and lower toxicity than “AB‐AmdU”. Remarkably, the addition of POM‐AmdU to the water of zebrafish larvae enabled the biosynthesis of azide‐modified DNA throughout the body.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Uncontrolled Keywords:Organic Chemistry, Molecular Medicine, Biochemistry, Molecular Biology
Language:English
Date:17 September 2018
Deposited On:07 Mar 2019 07:58
Last Modified:07 Mar 2019 08:12
Publisher:Wiley-VCH Verlag
ISSN:1439-4227
OA Status:Closed
Publisher DOI:https://doi.org/10.1002/cbic.201800351
Project Information:
  • : FunderSNSF
  • : Grant ID205320_165949
  • : Project TitleLive-cell imaging of DNA conformation and metabolism
  • : FunderDr. Helmut Legerlotz‐Stiftung
  • : Grant ID
  • : Project Title

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