Brightly Fluorescent 2'-Deoxyribonucleoside Triphosphates Bearing Methylated Bodipy Fluorophore for in Cellulo Incorporation to DNA, Imaging, and Flow Cytometry.
Fiche publication
Date publication
novembre 2018
Journal
Bioconjugate chemistry
Auteurs
Membres identifiés du Cancéropôle Est :
Dr DZIUBA Dmytro
Tous les auteurs :
Güixens-Gallardo P, Zawada Z, Matyašovský J, Dziuba D, Pohl R, Kraus T, Hocek M
Lien Pubmed
Résumé
Synthesis of cytosine, uracil, and 7-deazaadenine 2'-deoxyribonucleosides and triphosphates (dNTPs) bearing hexamethylated phenyl-bodipy fluorophore attached at position 5 of pyrimidines or at position 7 of 7-deazapurine was developed. All the title labeled nucleosides and dNTPs displayed bright green fluorescence with very high quantum yields. The modified dNTPs were good substrates to diverse DNA polymerases and were used for in vitro enzymatic synthesis of labeled DNA by primer extension or PCR. In combination with cationic cyclodextrin-peptide-based dNTP transporter, the dNTPs were successfully used for staining of genomic DNA in living cells for applications in confocal microscopy and in flow cytometry. The best performing cytosine nucleotide dCTP was used to monitor mitosis in live cells.
Mots clés
Adenine, analogs & derivatives, Boron Compounds, chemistry, Cell Line, Tumor, Cytosine, chemistry, DNA, analysis, Deoxyribonucleosides, chemistry, Flow Cytometry, Fluorescent Dyes, chemistry, Humans, Methylation, Optical Imaging, Polyphosphates, chemistry, Purines, chemistry, Staining and Labeling
Référence
Bioconjug Chem. 2018 11 21;29(11):3906-3912