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

• Published in: Organic & biomolecular chemistry Vol. 13; no. 15; pp. 4589 - 4595
• Main Authors: Seo, Siyoong, Onizuka, Kazumitsu, Nishioka, Chieko, Takahashi, Eiki, Tsuneda, Satoshi, Abe, Hiroshi, Ito, Yoshihiro
• Format: Journal Article
• Language: English
• Published: CAMBRIDGE Royal Soc Chemistry 21.04.2015
• Subjects: 3T3 Cells; Animals; Chemistry; Chemistry, Organic; Deoxyuridine - administration & dosage; Deoxyuridine - analogs & derivatives; Deoxyuridine - chemistry; Deoxyuridine - metabolism; DNA - analysis; DNA - metabolism; HeLa Cells; Humans; Mice; Phosphorylation; Physical Sciences; Science & Technology; Staining and Labeling - methods; CELLS; 5-ETHYNYLURACIL; AZIDES; PRODRUGS; IN-VIVO; EDU; TERMINAL ALKYNES
• ISSN: 1477-0520; 1477-0539
• DOI: 10.1039/c5ob00199d

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.