| Autor: |
Schreier VN; Department of Chemistry, University of Zurich, Zurich CH-8006, Switzerland., Loehr MO; Department of Chemistry, University of Zurich, Zurich CH-8006, Switzerland.; Department of Chemistry, McGill University, Montreal, Quebec H3A 0G4, Canada., Deng T; Department of Molecular Life Sciences, University of Zurich, Zurich CH-8006, Switzerland., Lattmann E; Department of Molecular Life Sciences, University of Zurich, Zurich CH-8006, Switzerland., Hajnal A; Department of Molecular Life Sciences, University of Zurich, Zurich CH-8006, Switzerland., Neuhauss SCF; Department of Molecular Life Sciences, University of Zurich, Zurich CH-8006, Switzerland., Luedtke NW; Department of Chemistry, University of Zurich, Zurich CH-8006, Switzerland.; Department of Chemistry, McGill University, Montreal, Quebec H3A 0G4, Canada.; Department of Pharmacology and Therapeutics, McGill University, Montreal, Quebec H3A 0G4, Canada. |
| Abstrakt: |
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 . |
