Abstract
Viruses are among the simplest biological systems and are highly effective vehicles for the delivery of genetic material into susceptible host cells1. Artificial viruses can be used as model systems for providing insights into natural viruses and can be considered a testing ground for developing artificial life. Moreover, they are used in biomedical and biotechnological applications, such as targeted delivery of nucleic acids for gene therapy1,2 and as scaffolds in material science3,4,5. In a natural setting, survival of viruses requires that a significant fraction of the replicated genomes be completely protected by coat proteins. Complete protection of the genome is ensured by a highly cooperative supramolecular process between the coat proteins and the nucleic acids, which is based on reversible, weak and allosteric interactions only6,7,8,9. However, incorporating this type of supramolecular cooperativity into artificial viruses remains challenging10,11,12,13,14,15. Here, we report a rational design for a self-assembling minimal viral coat protein based on simple polypeptide domains. Our coat protein features precise control over the cooperativity of its self-assembly with single DNA molecules to finally form rod-shaped virus-like particles. We confirm the validity of our design principles by showing that the kinetics of self-assembly of our virus-like particles follows a previous model developed for tobacco mosaic virus9. We show that our virus-like particles protect DNA against enzymatic degradation and transfect cells with considerable efficiency, making them promising delivery vehicles.
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Acknowledgements
The authors thank A. Westphal and I. van Hees for help with the fluorescence correlation spectroscopy experiments and R. Fokkink for help with static light scattering. The authors also thank M.W.P. van de Put for help in performing part of the cryo-TEM experiments and W. Kegel for suggesting the electrophoretic light scattering experiment. A.H-G. is financially supported by the Dutch Polymer Institute (DPI), project #698 SynProt, and by the Consejo Nacional de Ciencia y Tecnología (CONACyT), México. M.E.F. is supported by the Dutch Polymer Institute (DPI, Technology area HTE, project #730). M.C.S. is supported by the European Research Council (advanced grant no. 267254). D.J.K. acknowledges financial support through a Rubicon fellowship (grant no. 680-50-1019) from the Netherlands Organization for Scientific Research (NWO).
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F.d.W., M.C.S., P.v.d.S. and R.d.V. conceived the initial idea. R.d.V. and A.H-G. conceived and designed the molecules and experiments. A.H-G. produced the proteins, except for C–S2–B, which was produced by A.F.J.J. M.W. contributed to the production of proteins and to writing the technical sections on protein production. Except for the transfection experiments, analytical ultracentrifugation and cryo-TEM imaging, A.H-G. performed all experiments. A.H-G. and R.d.V. interpreted and analysed all experimental data except for the transfection experiments. P.v.d.S. and D.J.K. performed the theoretical analysis of the VLP assembly kinetics. R.B. designed the transfection experiments. M.F. performed and analysed the transfection experiments. D.M.E.T-W. designed, performed and analysed the analytical ultracentrifugation experiments. N.A.J.M.S. designed the cryo-TEM imaging experiments. P.H.H.B. performed the cryo-TEM imaging. A.H-G., D.J.K., P.v.d.S., M.C.S. and R.d.V. wrote the paper. All authors commented on the manuscript.
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Hernandez-Garcia, A., Kraft, D., Janssen, A. et al. Design and self-assembly of simple coat proteins for artificial viruses. Nature Nanotech 9, 698–702 (2014). https://doi.org/10.1038/nnano.2014.169
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DOI: https://doi.org/10.1038/nnano.2014.169
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