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Cellular Uptake of Tile-Assembled DNA Nanotubes

DNA-based nanostructures have received great attention as molecular vehicles for cellular delivery of biomolecules and cancer drugs. Here, we report on the cellular uptake of tubule-like DNA tile-assembled nanostructures 27 nm in length and 8 nm in diameter that carry siRNA molecules, folic acid and...

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Autores principales: Kocabey, Samet, Meinl, Hanna, MacPherson, Iain S., Cassinelli, Valentina, Manetto, Antonio, Rothenfusser, Simon, Liedl, Tim, Lichtenegger, Felix S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5312849/
https://www.ncbi.nlm.nih.gov/pubmed/28346998
http://dx.doi.org/10.3390/nano5010047
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author Kocabey, Samet
Meinl, Hanna
MacPherson, Iain S.
Cassinelli, Valentina
Manetto, Antonio
Rothenfusser, Simon
Liedl, Tim
Lichtenegger, Felix S.
author_facet Kocabey, Samet
Meinl, Hanna
MacPherson, Iain S.
Cassinelli, Valentina
Manetto, Antonio
Rothenfusser, Simon
Liedl, Tim
Lichtenegger, Felix S.
author_sort Kocabey, Samet
collection PubMed
description DNA-based nanostructures have received great attention as molecular vehicles for cellular delivery of biomolecules and cancer drugs. Here, we report on the cellular uptake of tubule-like DNA tile-assembled nanostructures 27 nm in length and 8 nm in diameter that carry siRNA molecules, folic acid and fluorescent dyes. In our observations, the DNA structures are delivered to the endosome and do not reach the cytosol of the GFP-expressing HeLa cells that were used in the experiments. Consistent with this observation, no elevated silencing of the GFP gene could be detected. Furthermore, the presence of up to six molecules of folic acid on the carrier surface did not alter the uptake behavior and gene silencing. We further observed several challenges that have to be considered when performing in vitro and in vivo experiments with DNA structures: (i) DNA tile tubes consisting of 42 nt-long oligonucleotides and carrying single- or double-stranded extensions degrade within one hour in cell medium at 37 °C, while the same tubes without extensions are stable for up to eight hours. The degradation is caused mainly by the low concentration of divalent ions in the media. The lifetime in cell medium can be increased drastically by employing DNA tiles that are 84 nt long. (ii) Dyes may get cleaved from the oligonucleotides and then accumulate inside the cell close to the mitochondria, which can lead to misinterpretation of data generated by flow cytometry and fluorescence microscopy. (iii) Single-stranded DNA carrying fluorescent dyes are internalized at similar levels as the DNA tile-assembled tubes used here.
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spelling pubmed-53128492017-03-21 Cellular Uptake of Tile-Assembled DNA Nanotubes Kocabey, Samet Meinl, Hanna MacPherson, Iain S. Cassinelli, Valentina Manetto, Antonio Rothenfusser, Simon Liedl, Tim Lichtenegger, Felix S. Nanomaterials (Basel) Article DNA-based nanostructures have received great attention as molecular vehicles for cellular delivery of biomolecules and cancer drugs. Here, we report on the cellular uptake of tubule-like DNA tile-assembled nanostructures 27 nm in length and 8 nm in diameter that carry siRNA molecules, folic acid and fluorescent dyes. In our observations, the DNA structures are delivered to the endosome and do not reach the cytosol of the GFP-expressing HeLa cells that were used in the experiments. Consistent with this observation, no elevated silencing of the GFP gene could be detected. Furthermore, the presence of up to six molecules of folic acid on the carrier surface did not alter the uptake behavior and gene silencing. We further observed several challenges that have to be considered when performing in vitro and in vivo experiments with DNA structures: (i) DNA tile tubes consisting of 42 nt-long oligonucleotides and carrying single- or double-stranded extensions degrade within one hour in cell medium at 37 °C, while the same tubes without extensions are stable for up to eight hours. The degradation is caused mainly by the low concentration of divalent ions in the media. The lifetime in cell medium can be increased drastically by employing DNA tiles that are 84 nt long. (ii) Dyes may get cleaved from the oligonucleotides and then accumulate inside the cell close to the mitochondria, which can lead to misinterpretation of data generated by flow cytometry and fluorescence microscopy. (iii) Single-stranded DNA carrying fluorescent dyes are internalized at similar levels as the DNA tile-assembled tubes used here. MDPI 2014-12-30 /pmc/articles/PMC5312849/ /pubmed/28346998 http://dx.doi.org/10.3390/nano5010047 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kocabey, Samet
Meinl, Hanna
MacPherson, Iain S.
Cassinelli, Valentina
Manetto, Antonio
Rothenfusser, Simon
Liedl, Tim
Lichtenegger, Felix S.
Cellular Uptake of Tile-Assembled DNA Nanotubes
title Cellular Uptake of Tile-Assembled DNA Nanotubes
title_full Cellular Uptake of Tile-Assembled DNA Nanotubes
title_fullStr Cellular Uptake of Tile-Assembled DNA Nanotubes
title_full_unstemmed Cellular Uptake of Tile-Assembled DNA Nanotubes
title_short Cellular Uptake of Tile-Assembled DNA Nanotubes
title_sort cellular uptake of tile-assembled dna nanotubes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5312849/
https://www.ncbi.nlm.nih.gov/pubmed/28346998
http://dx.doi.org/10.3390/nano5010047
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