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Subcellular-resolution delivery of a cytokine through precisely manipulated nanowires

Abstract

Precise delivery of molecular doses of biologically active chemicals to a pre-specified single cell among many, or a specific subcellular location, is still a largely unmet challenge hampering our understanding of cell biology. Overcoming this could allow unprecedented levels of cell manipulation and targeted intervention. Here, we show that gold nanowires conjugated with a cytokine such as tumour-necrosis factor-alpha can be transported along any prescribed trajectory or orientation using electrophoretic and dielectrophoretic forces to a specific location with subcellular resolution. The nanowire, 6 µm long and 300 nm in diameter, delivered the cytokine and activated canonical nuclear factor-kappaB signalling in a single cell. Combined computational modelling and experimentation indicated that cell stimulation was highly localized to the nanowire vicinity. This targeted delivery method has profound implications for controlling signalling events on the single cell level.

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Figure 1: Nanowire functionalization and movement.
Figure 2: Nanowire delivery to pre-selected cells.
Figure 3: Delivery of functional TNFα by nanowires.
Figure 4: Simulations predict localized TNFα delivery.
Figure 5: Selective stimulation by TNFα-coated nanowires.

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Acknowledgements

The authors thank W. Greene of UCSF for providing the p65 plasmid, J. Wang for helping with cell transfection, X. Li for TNFα labelling and Z. Wang for microplate scanning. D.F., F.Q.Z., C.L.C. acknowledge support from the National Science Foundation (DMR 0403849). R.C.C. acknowledges support from the National Science Foundation (DMR 0706178). Z.Y., R.C. and A.L. acknowledge the support from the National Institutes of Health (GM072024, RR020839). D.F. acknowledges start up support from University of Texas at Austin. R.C. acknowledges support from the Medical Scientist Training Program at Johns Hopkins University.

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D.F., Z.Y., C.L.C. and A.L. conceived and designed the experiments and analysis. Z.Y., D.F. and F.Q.Z. performed the experiments and analysed the data. R.C. performed the simulations. All authors discussed the results and co-wrote the paper.

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Correspondence to C. L. Chien or Andre Levchenko.

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The authors declare no competing financial interests.

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Fan, D., Yin, Z., Cheong, R. et al. Subcellular-resolution delivery of a cytokine through precisely manipulated nanowires. Nature Nanotech 5, 545–551 (2010). https://doi.org/10.1038/nnano.2010.104

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