Brightly Fluorescent 2'-Deoxyribonucleoside Triphosphates Bearing Methylated Bodipy Fluorophore for in Cellulo Incorporation to DNA, Imaging, and Flow Cytometry

0498658 - ÚOCHB 2019 RIV US eng J - Článek v odborném periodiku
Güixens-Gallardo, Pedro - Zawada, Zbigniew - Matyašovský, Ján - Dziuba, Dmytro - Pohl, Radek - Kraus, Tomáš - Hocek, Michal
Brightly Fluorescent 2'-Deoxyribonucleoside Triphosphates Bearing Methylated Bodipy Fluorophore for in Cellulo Incorporation to DNA, Imaging, and Flow Cytometry.
Bioconjugate Chemistry. Roč. 29, č. 11 (2018), s. 3906-3912. ISSN 1043-1802. E-ISSN 1520-4812
Grant CEP: GA ČR GBP206/12/G151; GA ČR(CZ) GA17-14791S; GA MŠMT(CZ) EF16_019/0000729
Grant ostatní: AV ČR(CZ) AP1501
Program: Akademická prémie - Praemium Academiae
Institucionální podpora: RVO:61388963
Klíčová slova: nucleoside triphosphates * labeled nucleotides * enzymatic synthesis
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 4.349, rok: 2018
Způsob publikování: Open access
https://pubs.acs.org/doi/10.1021/acs.bioconjchem.8b00721

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 dN(mBdp)TPs 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 dN(mBdp)TPs 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 dN(mBdp)TPs was used to monitor mitosis in live cells.
Trvalý link: http://hdl.handle.net/11104/0290953