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Labeling of Cellular DNA with a Cyclo sal Phosphotriester Pronucleotide Analog of 5‐ethynyl‐2′‐deoxyuridine
Author(s) -
Huynh Ngoc,
Dickson Charlotte,
Zencak Dusan,
Hilko David H.,
MackaySim Alan,
Poulsen SallyAnn
Publication year - 2015
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/cbdd.12506
Subject(s) - bioconjugation , deoxyuridine , chemistry , dna , chemical biology , nucleoside , biochemistry , azide , click chemistry , small molecule , nucleic acid , combinatorial chemistry , organic chemistry
DNA synthesis is a fundamental biological process central to all proliferating cells, and the design of small molecule probes that allow detection of this DNA is important for many applications. 5‐Ethynyl‐2′‐deoxyuridine, known as EdU, has become a workhorse for metabolic labeling of DNA in mammalian cells, followed by bioconjugation to a small molecule fluorescent azide using copper‐catalyzed azide–alkyne cycloaddition (Cu AAC ), click chemistry, to allow detection. In this study, we demonstrate that a cyclo sal phosphotriester pronucleotide analog of EdU is suitable for metabolic incorporation into DNA of proliferating cells and subsequent labeling by Cu AAC . This analog has two advantages over EdU; first, by delivering EdU with a preinstalled 5′‐monophosphate moiety, it bypasses the need for thymidine kinase processing, and second, the increased lipophilicity compared to EdU may enable passive diffusion across the cell membrane and may circumvent the reliance on nucleoside active transport mechanisms for cellular uptake. These advantages pave the way for the development of additional novel pronucleotides to widen experimental opportunities for future bioconjugation applications involving cellular DNA .