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Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles

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Abstract

The therapeutic promise of small interfering RNAs (siRNAs) for specific gene silencing is dependent on the successful delivery of functional siRNAs to the cytoplasm. Their conjugation to an established delivery platform, such as gold nanoparticles, offers tremendous potential for treating diseases and advancing our understanding of cellular processes. Their success or failure is dependent on both the uptake of the nanoparticles into the cells and subsequent intracellular release of the functional siRNA. In this study, utilizing gold nanoparticle siRNA-mediated delivery against C-MYC, we aimed to determine if we could achieve knockdown in a cancer cell line with low levels of intracellular glutathione, and determine the influence, if any, of polyethylene glycol (PEG) ligand density on knockdown, with a view to determining the optimal nanoparticle design to achieve C-MYC knockdown. We demonstrate that, regardless of the PEG density, knockdown in cells with relatively low glutathione levels can be achieved, as well as the possible effect of steric hindrance of PEG on the availability of the siRNA for cleavage in the intracellular environment. Gold nanoparticle uptake was demonstrated via transmission electron microscopy and mass spectroscopy, while knockdown was determined at the protein and physiological levels (cells in S-phase) by in-cell westerns and BrdU incorporation, respectively.

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Authors and Affiliations

  1. Centre for Cell Engineering, University of Glasgow, Glasgow, G12 8QQ, Scotland, UK

    Mark McCully & Catherine C. Berry

  2. Institute of Nanoscience of Aragon, University of Zaragoza, Zaragoza, 50018, Spain

    Yulan Hernandez & Jesus M. de la Fuente

  3. Massachusetts Institute of Technology, Institute for Medical Engineering and Science, Harvard-MIT Division for Health Sciences and Technology, Cambridge, Massachusetts, 02139, USA

    João Conde

  4. UCIBIO, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, Caparica, 2829-516, Portugal

    Pedro V. Baptista

  5. Instituto de Ciencia de Materiales de Aragón (ICMA), CSIC-Universidad de Zaragoza, c/Pedro Cerbuna 12, Zaragoza, 50009, Spain

    Jesus M. de la Fuente

  6. Institute of Nano Biomedicine and Engineering, Key Laboratory for Thin Film and Microfabrication Technology of the Ministry of Education, Research Institute of Translation Medicine, Shanghai Jiao Tong University, Dongchuan Road 800, Shanghai, 200240, China

    Jesus M. de la Fuente

  7. School of Science, University of the West of Scotland, Paisley, PA1 2BE, Scotland, UK

    Andrew Hursthouse & David Stirling

Authors
  1. Mark McCully
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  2. Yulan Hernandez
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  3. João Conde
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  4. Pedro V. Baptista
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  5. Jesus M. de la Fuente
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Correspondence to Mark McCully.

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McCully, M., Hernandez, Y., Conde, J. et al. Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles. Nano Res. 8, 3281–3292 (2015). https://doi.org/10.1007/s12274-015-0828-5

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  • DOI: https://doi.org/10.1007/s12274-015-0828-5

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