Abstract
A modified 2'-deoxycytidine triphosphate derivative (dCTOTP) bearing thiazole orange moiety tethered via an oligoethylene glycol linker was designed and synthesized. The nucleotide was found to incorporate into DNA by DNA polymerases in vitro as well as in live cells. Upon incorporation of dCTOTP 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 dCTOTP. When dCTOTP was delivered into live U-2 OS cells using a synthetic nucleoside triphosphate transporter, it allowed us to distinguish and monitor in real time the cells that were actively synthesizing DNA from the very first moments after treatment. We anticipate that this probe can be used to study chromatin organization and dynamics.
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