Modifikované nukleosid trifosfáty s připojenými fluorofory pro metabolické značení DNA

Hájek, Tomáš

Fluorescent labeling techniques help us understand how DNA replication determines cell division, DNA repair, and disease progression at a molecular level by visualizing DNA synthesis. However, a shortage of nucleotide triphosphates bearing fluorophores that operate in the long-wavelength region of the visible spectrum remains. In this work, nucleoside triphosphates tagged with a fluorescent label emitting in the red region of the spectrum for metabolic labeling were developed. The three prepared nucleotides carry a thiazole orange- derived fluorophore, which intercalates into double-stranded DNA upon incorporation. Notably, one of the compounds also produces a distinct signal at different excitation wavelengths in mitochondria, suggesting its potential as a dual-function probe for simultaneous visualization of nuclear replication and mitochondrial distribution within the same cell. This approach expands the toolkit for studying DNA replication dynamics in live cells and may advance both imaging methodologies and disease diagnostics.

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