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Fluorescence resonance energy transfer between green fluorescent protein and doxorubicin enabled by DNA nanotechnology

Heger, Zbynek, Kominkova, Marketa, Cernei, Natalia, Krejcova, Ludmila, Kopel, Pavel, Zitka, Ondrej, Adam, Vojtech, Kizek, Rene
Electrophoresis 2014 v.35 no.23 pp. 3290-3301
DNA, doxorubicin, electrophoresis, energy transfer, fluorescence, fullerene, gold, green fluorescent protein, nanogold, thermotherapy, thiols, wavelengths
DNA nanotechnology is a rapidly growing research area, where DNA may be used for wide range of applications such as construction of nanodevices serving for large scale of diverse purposes. Likewise a panel of various purified fluorescent proteins is investigated for their ability to emit their typical fluorescence spectra under influence of particular excitation. Hence these proteins may form ideal donor molecules for assembly of fluorescence resonance emission transfer (FRET) constructions. To extend the application possibilities of fluorescent proteins, while using DNA nanotechnology, we developed nanoconstruction comprising green fluorescent protein (GFP) bound onto surface of surface active nanomaghemite and functionalized with gold nanoparticles. We took advantage of natural affinity between gold and thiol moieties, which were modified to bind DNA fragment. Finally we enclosed doxorubicin into fullerene cages. Doxorubicin intercalated in DNA fragment bound on the particles and thus we were able to connect these parts together. Because GFP behaved as a donor and doxorubicin as an acceptor using excitation wavelength for GFP (395 nm) in emission wavelength of doxorubicin (590 nm) FRET was observed. This nanoconstruction may serve as a double‐labeled transporter of doxorubicin guided by force of external magnetic force owing to the presence of nanomaghemite. Further nanomaghemite offers the possibility of using this technology for thermotherapy.