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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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.
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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.
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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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DOI: https://doi.org/10.1038/nchem.2472
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