PUBLICATION
Fluorescent dATP for DNA Synthesis In Vivo
- Authors
- Schreier, V.N., Loehr, M.O., Deng, T., Lattmann, E., Hajnal, A., Neuhauss, S.C.F., Luedtke, N.W.
- ID
- ZDB-PUB-201029-1
- Date
- 2020
- Source
- ACS Chemical Biology 15(11): 2996-3003 (Journal)
- Registered Authors
- Neuhauss, Stephan
- Keywords
- none
- MeSH Terms
-
- Animals
- Boron Compounds/chemical synthesis
- Boron Compounds/chemistry
- Caenorhabditis elegans/ultrastructure
- Carbocyanines/chemical synthesis
- Carbocyanines/chemistry
- DNA/analysis*
- DNA Replication*
- Deoxyadenine Nucleotides/chemical synthesis
- Deoxyadenine Nucleotides/chemistry*
- Fluorescent Dyes/chemical synthesis
- Fluorescent Dyes/chemistry*
- Optical Imaging/methods
- Rhodamines/chemical synthesis
- Rhodamines/chemistry
- Zebrafish/embryology
- PubMed
- 33108866 Full text @ ACS Chem. Biol.
Citation
Schreier, V.N., Loehr, M.O., Deng, T., Lattmann, E., Hajnal, A., Neuhauss, S.C.F., Luedtke, N.W. (2020) Fluorescent dATP for DNA Synthesis In Vivo. ACS Chemical Biology. 15(11):2996-3003.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping