Work

Fluorescent dATP for DNA Synthesis In Vivo

Year: 2020

Type: article

Abstract: Fluorescent nucleoside triphosphates are powerful probes of DNA synthesis, but their potential use in living animals has been previously underexplored. Here, we report the synthesis and characterization of 7-deaza-(1,2,3-triazole)-2′-deoxyadenosine-5′-triphosphate (dATP) derivatives of tetramethyl rhodamine (“TAMRA-dATP”), cyanine (“Cy3-dATP”), and boron-dipyrromethene (“BODIPY-dATP”). Upon microinjection into live zebrafish embryos, all three compounds were incorporated into the DNA of dividing cells; however, their impact on embryonic toxicity was highly variable, depending on the exact structure of the dye. TAMRA-EdATP exhibited superior characteristics in terms of its high brightness, low toxicity, and rapid incorporation and depletion kinetics in both a vertebrate (zebrafish) and a nematode ( Caenorhabditis elegans ). TAMRA-EdATP allows for unprecedented, real-time visualization of DNA replication and chromosome segregation in vivo .

Source: ACS Chemical Biology

DOI: 10.1021/acschembio.0c00654