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Self-assembly of size-controlled liposomes on DNA nanotemplates

Nature Chemistry volume 8, pages 476–483 (2016)Cite this article

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Abstract

Artificial lipid-bilayer membranes are valuable tools for the study of membrane structure and dynamics. For applications such as the study of vesicular transport and drug delivery, there is a pressing need for artificial vesicles with controlled size. However, controlling vesicle size and shape with nanometre precision is challenging, and approaches to achieve this can be heavily affected by lipid composition. Here, we present a bio-inspired templating method to generate highly monodispersed sub-100-nm unilamellar vesicles, where liposome self-assembly was nucleated and confined inside rigid DNA nanotemplates. Using this method, we produce homogeneous liposomes with four distinct predefined sizes. We also show that the method can be used with a variety of lipid compositions and probe the mechanism of templated liposome formation by capturing key intermediates during membrane self-assembly. The DNA nanotemplating strategy represents a conceptually novel way to guide lipid bilayer formation and could be generalized to engineer complex membrane/protein structures with nanoscale precision.

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Figure 1: Scheme for generating size-controlled vesicles by nanotemplating.
Figure 2: Characterization of DNA-templated liposomes.
Figure 3: Effect of initial lipid seed placement on final vesicle formation.
Figure 4: Vesicle formation mechanism studied by capturing intermediates during dialysis.

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Acknowledgements

The authors thank P.D. Ellis for designing the DNA rings used for placing lipid seeds at different angles and for proofreading the manuscript, and F. Sigworth for providing cryo-electron micrographs of extruded liposomes. This work is supported by a National Institutes of Health (NIH) Director's New Innovator Award (DP2-GM114830), an NIH grant (R21-GM109466) and a Yale University faculty startup fund to C.L., an NIH grant to J.E.R. (R01-DK027044) and an NIH Director's New Innovator Award (DP2-OD004641), an Army Research Office MURI grant (W911NF-12-1-0420), National Science Foundation Expeditions Grant (1317694) and a Wyss Institute for Biologically Inspired Engineering Faculty Award to W.M.S.

Author information

Author notes

  1. Hideki Shigematsu

    Present address: Present address: Division of Structural and Synthetic Biology, RIKEN Center for Life Science Technologies, Yokohama, Kangawa 230-0045, Japan,

  2. Yang Yang and Jing Wang: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Cell Biology, Yale University School of Medicine, New Haven, 06520, Connecticut, USA

    Yang Yang, Jing Wang, Weiming Xu, James E. Rothman & Chenxiang Lin

  2. Nanobiology Institute, Yale University, West Haven, 06516, Connecticut, USA

    Yang Yang, Jing Wang, Weiming Xu, James E. Rothman & Chenxiang Lin

  3. Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, 06250, Connecticut, USA

    Hideki Shigematsu

  4. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, 02138, Massachusetts, USA

    William M. Shih

  5. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    William M. Shih

  6. Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, 02215, Massachusetts, USA

    William M. Shih

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Contributions

Y.Y. and J.W. designed and conducted the majority of the experiments, analysed the data, prepared the majority of the manuscript and contributed equally to this work. H.S. performed cryo-EM studies and prepared the manuscript. W.X. initiated the project, developed the DNA–lipid conjugation method and designed and performed pilot experiments to prepare and purify DNA-ring enclosed vesicles. W.M.S. initiated the project and discussed the results. J.E.R. initiated the project, supervised J.W. and W.X., and discussed the results. C.L. initiated the project, designed and supervised the study, interpreted the data and prepared the manuscript. All authors reviewed and approved the manuscript.

Corresponding authors

Correspondence to William M. Shih, James E. Rothman or Chenxiang Lin.

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The authors declare no competing financial interests.

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Yang, Y., Wang, J., Shigematsu, H. et al. Self-assembly of size-controlled liposomes on DNA nanotemplates. Nature Chem 8, 476–483 (2016). https://doi.org/10.1038/nchem.2472

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