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Cell-free protein synthesis and assembly on a biochip

Nature Nanotechnology volume 7, pages 374–378 (2012)Cite this article

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Abstract

Biologically active complexes such as ribosomes and bacteriophages are formed through the self-assembly of proteins and nucleic acids1,2. Recapitulating these biological self-assembly processes in a cell-free environment offers a way to develop synthetic biodevices3,4,5,6. To visualize and understand the assembly process, a platform is required that enables simultaneous synthesis, assembly and imaging at the nanoscale. Here, we show that a silicon dioxide grid, used to support samples in transmission electron microscopy, can be modified into a biochip to combine in situ protein synthesis, assembly and imaging. Light is used to pattern the biochip surface with genes that encode specific proteins, and antibody traps that bind and assemble the nascent proteins. Using transmission electron microscopy imaging we show that protein nanotubes synthesized on the biochip surface in the presence of antibody traps efficiently assembled on these traps, but pre-assembled nanotubes were not effectively captured. Moreover, synthesis of green fluorescent protein from its immobilized gene generated a gradient of captured proteins decreasing in concentration away from the gene source. This biochip could be used to create spatial patterns of proteins assembled on surfaces.

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Figure 1: Schematic showing the TEM biochip.
Figure 2: Correlating patterns of GFP fluorescence with TEM imaging of gold-labelled antibodies.
Figure 3: Synthesis, trapping and assembly of nanotube-forming proteins.
Figure 4: TEM imaging of DNA revealed by RecA protein polymerization.
Figure 5: Combined synthesis, trapping and gradients of fluorescent proteins on the chip.

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Acknowledgements

This work was supported by the Israel Science Foundation (Converging Technology programme) and the Minerva Foundation. The electron microscopy studies were conducted at the Irving and Cherna Moskowitz Center for Nano and Bio-Nano Imaging at the Weizmann Institute of Science. The authors thank T. Arad and V. Shinder for TEM assistance.

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Author notes

  1. Amnon Buxboim

    Present address: Present address: Department of Physics and Astronomy, University of Pennsylvania, Philadelphia, Pennsylvania 19103, USA,

Authors and Affiliations

  1. Department of Materials and Interfaces, The Weizmann Institute of Science, Rehovot, 76100, Israel

    Yael Heyman, Amnon Buxboim, Shirley S. Daube & Roy H. Bar-Ziv

  2. Chemical Research Support, The Weizmann Institute of Science, Rehovot, 76100, Israel

    Sharon G. Wolf

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  1. Yael Heyman
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Contributions

Y.H., S.S.D. and R.H.B-Z. conceived and designed the experiments. Y.H., A.B. and S.S.D. performed the experiments. S.G.W. contributed TEM expertise and materials/analysis tools. Y.H., S.S.D. and R.H.B-Z. co-wrote the paper. All authors analysed the data, discussed the results and commented on the manuscript.

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Correspondence to Roy H. Bar-Ziv.

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The authors declare no competing financial interests.

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Heyman, Y., Buxboim, A., Wolf, S. et al. Cell-free protein synthesis and assembly on a biochip. Nature Nanotech 7, 374–378 (2012). https://doi.org/10.1038/nnano.2012.65

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