Phosphorylated 5-ethynyl-2′-deoxyuridine for advanced DNA labeling

Siyoong Seo, Kazumitsu Onizuka, Chieko Nishioka, Eiki Takahashi, Satoshi Tsuneda, Hiroshi Abe, Yoshihiro Ito

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    Abstract

    The representative DNA-labeling agent 5-ethynyl-2′-deoxyuridine (EdU) was chemically modified to improve its function. Chemical monophosphorylation was expected to enhance the efficiency of the substrate in DNA polymerization by circumventing the enzymatic monophosphorylation step that consumes energy. In addition, to enhance cell permeability, the phosphates were protected with bis-pivaloyloxymethyl that is stable in buffer and plasma, and degradable inside various cell types. The phosphorylated EdU (PEdU) was less toxic than EdU, and had the same or a slightly higher DNA-labeling ability in vitro. PEdU was also successfully applied to DNA labeling in vivo. In conclusion, PEdU can be used as a less toxic DNA-labeling agent for studies that require long-term cell survival or very sensitive cell lines.

    Original languageEnglish
    Pages (from-to)4589-4595
    Number of pages7
    JournalOrganic and Biomolecular Chemistry
    Volume13
    Issue number15
    DOIs
    Publication statusPublished - 2015 Apr 21

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    ASJC Scopus subject areas

    • Physical and Theoretical Chemistry
    • Organic Chemistry
    • Biochemistry

    Cite this

    Seo, S., Onizuka, K., Nishioka, C., Takahashi, E., Tsuneda, S., Abe, H., & Ito, Y. (2015). Phosphorylated 5-ethynyl-2′-deoxyuridine for advanced DNA labeling. Organic and Biomolecular Chemistry, 13(15), 4589-4595. https://doi.org/10.1039/c5ob00199d