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 - Journal Article
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
R&D Projects: GA ČR GBP206/12/G151; GA ČR(CZ) GA17-14791S; GA MŠMT(CZ) EF16_019/0000729
Grant - others:AV ČR(CZ) AP1501
Program: Akademická prémie - Praemium Academiae
Institutional support: RVO:61388963
Keywords : nucleoside triphosphates * labeled nucleotides * enzymatic synthesis
OECD category: Biochemistry and molecular biology
Impact factor: 4.349, year: 2018
Method of publishing: 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.
Permanent Link: http://hdl.handle.net/11104/0290953