PUBLICATION
Incorporation of azide groups into DNA using membrane-permeable nucleotide triesters in vivo
- Authors
- Tera, M., Glasauer, S., Luedtke, N.W.
- ID
- ZDB-PUB-180629-8
- Date
- 2018
- Source
- Chembiochem : a European journal of chemical biology 19(18): 1939-1943 (Journal)
- Registered Authors
- Glasauer, Stella
- Keywords
- DNA, chemical biology, click chemistry, metabolic labeling, nucleosides and nucleotides
- MeSH Terms
-
- Animals
- Azides/chemistry*
- Azides/metabolism
- Cell Membrane Permeability
- Click Chemistry
- DNA/chemistry*
- DNA/metabolism
- Esters/chemistry
- Esters/metabolism
- HeLa Cells
- Humans
- Nucleotides/chemistry*
- Nucleotides/metabolism
- Zebrafish
- Zidovudine/analogs & derivatives*
- Zidovudine/chemistry
- Zidovudine/metabolism
- PubMed
- 29953711 Full text @ Chembiochem
Citation
Tera, M., Glasauer, S., Luedtke, N.W. (2018) Incorporation of azide groups into DNA using membrane-permeable nucleotide triesters in vivo. Chembiochem : a European journal of chemical biology. 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 5-(azidomethyl)-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 carrying either 5'-bispivaloyloxymethyl (POM), 5'-bisacetoxybenzyl (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 superior 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping