Cellular uptake of ribonuclease A-functionalised core-shell silica microspheres


Analysis of protein function in a cellular context ideally requires physiologically representative levels of that protein. Thus conventional nucleic acid-based transfection methods are far from ideal owing to the over expression that generally results. Likewise fusions with protein transduction domains can be problematic whilst delivery via liposomes/nanoparticles typically results in endosomal localisation. Recently polymer microspheres have been reported to be highly effective at delivering proteins into cells and thus provide a viable new alternative for protein delivery (protein transduction). Herein we describe the successful delivery of active ribonuclease A into HeLa cells via novel polymer core-silica shell microspheres. Specifically, poly(styrene-co-vinylbenzylisothiouronium chloride) core particles, generated by dispersion polymerisation, were coated with a poly(styrene-co-trimethoxysilylpropyl methacrylate) shell. The resultant core-shell morphology was characterised by transmission electron, scanning electron and fluorescence confocal microscopies, whilst size and surface charge was assessed by dynamic light scattering and zeta-potential measurements, respectively. Subsequently ribonuclease A was coupled to the microspheres using simple carbodiimide chemistry. Gel electrophoresis confirmed and quantified the activity of the immobilised enzyme against purified HeLa RNA. Finally, the polymer-protein particles were evaluated as protein-transduction vectors in vitro to deliver active ribonuclease A to HeLa cells. Cellular uptake of the microspheres was successful and resulted in reduced levels of both intracellular RNA and cell viability.

Publication DOI: https://doi.org/10.1039/C4TB01130A
Divisions: College of Engineering & Physical Sciences > School of Infrastructure and Sustainable Engineering > Chemical Engineering & Applied Chemistry
College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences > Chronic and Communicable Conditions
College of Health & Life Sciences > School of Biosciences > Cell & Tissue Biomedical Research
College of Engineering & Physical Sciences > Aston Institute of Materials Research (AIMR)
Additional Information: This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Funding: EPSRC Industrial CASE (GFC) grant number EP/D038057/1 Electronic supplementary information: see DOI: 10.1039/c4tb01130a
Uncontrolled Keywords: Biomedical Engineering,Medicine(all),Chemistry(all),Materials Science(all)
Publication ISSN: 2050-750X
Last Modified: 17 Jun 2024 07:14
Date Deposited: 19 Aug 2019 10:06
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
http://pubs.rsc ... 0a#!divAbstract (Publisher URL)
PURE Output Type: Article
Published Date: 2014-11-14
Published Online Date: 2014-08-27
Authors: Chimonides, G.F
Behrendt, J.M.
Chundoo, E.
Bland, C.
Hine, A.V. (ORCID Profile 0000-0003-4065-831X)
Devitt, A. (ORCID Profile 0000-0002-4651-6761)
Nagel, D.A. (ORCID Profile 0000-0002-9055-1775)
Sutherland, A.J. (ORCID Profile 0000-0003-3651-1809)



Version: Published Version

License: Creative Commons Attribution

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