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Electron cryotomography (cryoET), an electron cryomicroscopy (cryoEM) modality, has changed our understanding of biological function by revealing the native molecular details of membranes, viruses, and cells. However, identification of individual molecules within tomograms from cryoET is challenging because of sample crowding and low signal-to-noise ratios. Here, we present a tagging strategy for cryoET that precisely identifies individual protein complexes in tomograms without relying on metal clusters. Our method makes use of DNA origami to produce "molecular signposts" that target molecules of interest, here via fluorescent fusion proteins, providing a platform generally applicable to biological surfaces. We demonstrate the specificity of signpost origami tags (SPOTs) in vitro as well as their suitability for cryoET of membrane vesicles, enveloped viruses, and the exterior of intact mammalian cells.

Original publication

DOI

10.1016/j.cell.2021.01.033

Type

Journal article

Journal

Cell

Publication Date

02/2021

Volume

184

Pages

1110 - 1121.e16

Addresses

Oxford Particle Imaging Centre, Division of Structural Biology, University of Oxford, Wellcome Centre for Human Genetics, Roosevelt Drive, Oxford UK OX3 7BN; Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford UK OX1 3PU; Centre for Structural Systems Biology, Heinrich-Pette-Institut, Leibniz-Institut für Experimentelle Virologie, Notkestrasse 85, 22607 Hamburg, Germany.

Keywords

Cell Line, Cell Membrane, Animals, Humans, DNA, Cryoelectron Microscopy, Fluorescence, Female, Aptamers, Nucleotide, Nanoparticles, Electron Microscope Tomography, Biophysical Phenomena