Uptake and Fate of Fluorescently Labeled DNA Nanostructures in Cellular Environments: A Cautionary Tale
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Uptake and Fate of Fluorescently Labeled DNA Nanostructures in Cellular Environments: A Cautionary Tale

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Uptake and Fate of Fluorescently Labeled DNA Nanostructures in Cellular Environments: A Cautionary Tale

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dc.contributor.author Lacroix, Aurélie
dc.contributor.author Vengut-Climent, Empar
dc.contributor.author de Rochambeau, Donatien
dc.contributor.author Sleiman, Hanadi F.
dc.date.accessioned 2019-07-17T10:08:40Z
dc.date.available 2019-07-17T10:08:40Z
dc.date.issued 2019 es_ES
dc.identifier.uri https://hdl.handle.net/10550/70855
dc.description.abstract Fluorescent dye labeling of DNA oligonucleotides and nanostructures is one of the most used techniques to track their fate and cellular localization inside cells. Here, we report that intracellular fluorescence, and even FRET signals, cannot be correlated with the cellular uptake of intact DNA structures. Live cell imaging revealed high colocalization of cyanine-labeled DNA oligos and nanostructures with phosphorylated small-molecule cyanine dyes, one of the degradation products from these DNA compounds. Nuclease degradation of the strands outside and inside the cell results in a misleading intracellular fluorescent signal. The signal is saturated by the fluorescence of the degradation product (phosphorylated dye). To test our hypothesis, we synthesized a range of DNA structures, including Cy3-and Cy5-labeled DNA cubes and DNA tetrahedra, and oligonucleotides with different stabilities toward nucleases. All give fluorescence signals within the mitochondria after cellular uptake and strongly colocalize with a free phosphorylated dye control. Kinetics experiments revealed that uptake of stable DNA structures is delayed. We also studied several parameters influencing fluorescent data: stability of the DNA strand, fixation methods that can wash away the signal, position of the dye on the DNA strand, and design of FRET experiments. DNA nanostructures hold tremendous potential for biomedical applications and biotechnology because of their biocompatibility, programmability, and easy synthesis. However, few examples of successful DNA machines in vivo have been reported. We believe this contribution can be used as a guide to design better cellular uptake experiments when using fluorescent dyes, in order to further propel the biological development, and application of DNA nanostructures. es_ES
dc.language.iso en_US es_ES
dc.source Lacroix, A.; Vengut-Climent, E.; de Rochambeau, D.; Sleiman, H. ACS Cent. Sci.2019, 5, 5, 882-891. ACS Publications es_ES
dc.subject cell uptake es_ES
dc.subject fluorescence es_ES
dc.subject dna nanostructures es_ES
dc.title Uptake and Fate of Fluorescently Labeled DNA Nanostructures in Cellular Environments: A Cautionary Tale es_ES
dc.type info:eu-repo/semantics/article es_ES
dc.subject.unesco UNESCO::QUÍMICA es_ES
dc.identifier.doi https://doi.org/10.1021/acscentsci.9b00174 es_ES

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