title: Phosphorylated 5-ethynyl-2'-deoxyuridine for advanced DNA labeling.
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.
authors.affiliation: Nano Medical Engineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. y-ito@riken.jp.
authors.forename: Siyoong
authors.initials: S
authors.lastname: Seo
authors.forename: Kazumitsu
authors.initials: K
authors.lastname: Onizuka
authors.forename: Chieko
authors.initials: C
authors.lastname: Nishioka
authors.forename: Eiki
authors.initials: E
authors.lastname: Takahashi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tsuneda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Abe
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ito
mesh_terms: D016475:3T3 Cells; D000818:Animals; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D003857:Deoxyuridine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010766:Phosphorylation; D013194:Staining and Labeling / Q000379:methods
chemical_list: D004247:DNA; C031086:5-ethynyl-2'-deoxyuridine; D003857:Deoxyuridine