Real-time Imaging of Nascent DNA in Live Cells by Monitoring the Fluorescence Lifetime of DNA-Incorporated Thiazole Orange-Modified Nucleotides

0574395 - ÚOCHB 2024 RIV DE eng J - Journal Article
Kuba, Miroslav - Khoroshyy, Petro - Lepšík, Martin - Kužmová, Erika - Kodr, David - Kraus, Tomáš - Hocek, Michal
Real-time Imaging of Nascent DNA in Live Cells by Monitoring the Fluorescence Lifetime of DNA-Incorporated Thiazole Orange-Modified Nucleotides.
Angewandte Chemie - International Edition. Roč. 62, č. 38 (2023), č. článku e202307548. ISSN 1433-7851. E-ISSN 1521-3773
R&D Projects: GA ČR(CZ) GX20-00885X
Institutional support: RVO:61388963
Keywords : DNA synthesis * fluorescence lifetime imaging microscopy * fluorophores * metabolic labelling * nucleotides
OECD category: Organic chemistry
Impact factor: 16.6, year: 2022
Method of publishing: Open access
https://doi.org/10.1002/anie.202307548

A modified 2′-deoxycytidine triphosphate derivative (dC(TO)TP) bearing a thiazole orange moiety tethered via an oligoethylene glycol linker was designed and synthesized. The nucleotide was incorporated into DNA by DNA polymerases in vitro as well as in live cells. Upon incorporation of dC(TO)TP into DNA, the thiazole orange moiety exhibited a fluorescence lifetime that differed significantly from the non-incorporated (i.e. free and non-covalently intercalated) forms of dC(TO)TP. When dC(TO)TP was delivered into live U-2 OS cells using a synthetic nucleoside triphosphate transporter, it allowed us to distinguish and monitor cells that were actively synthesizing DNA in real time, from the very first moments after the treatment. We anticipate that this probe could be used to study chromatin organization and dynamics.
Permanent Link: https://hdl.handle.net/11104/0344723