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DNA probes for monitoring dynamic and transient molecular encounters on live cell membranes

Abstract

Cells interact with the extracellular environment through molecules expressed on the membrane. Disruption of these membrane-bound interactions (or encounters) can result in disease progression. Advances in super-resolution microscopy have allowed membrane encounters to be examined, however, these methods cannot image entire membranes and cannot provide information on the dynamic interactions between membrane-bound molecules. Here, we show a novel DNA probe that can transduce transient membrane encounter events into readable cumulative fluorescence signals. The probe, which translocates from one anchor site to another, mimicking motor proteins, is realized through a toehold-mediated DNA strand displacement reaction. Using this probe, we successfully monitored rapid encounter events of membrane lipid domains using flow cytometry and fluorescence microscopy. Our results show a preference for encounters within the same lipid domains.

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Figure 1: Anchoring and operation scheme of DNA probe on live cell membrane.
Figure 2: Locomotion of DNA probe on live cell membrane.
Figure 3: DNA probe competition game to study encounter preference.
Figure 4: Locomotion of DNA probe on model lipid monolayer film.
Figure 5: DNA probe–aptamer conjugates to study membrane protein encounter rates.

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Acknowledgements

The authors thank X. Fang at Chinese Academy of Sciences, G. Fanucci, V. D. Kleiman and Y. Tseng at University of Florida, and K. Salaita at Emory University for helpful discussions and suggestions. This work is supported by NSFC grants (NSFC 21521063 and NSFC 21327009), and the US National Institutes of Health (GM079359 and CA133086). The work of G.B. was supported in part by a Key Project of the Major Research Plan of NSFC (no. 91130004), a NSFC A3 Project (no.11421110002), NSFC Tianyuan Projects (no. 11426235; no. 11526211), a NSFC Innovative Group Fund (no.11621101) and US NSF FRG DMS-0968360.

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Contributions

M.Y., Y.L. and D.H. contributed equally to this work. M.Y., D.H. and W.T. conceived and designed the experiments. M.Y. and Y.L. performed the experiments. M.Y., D.H., L.P. and G.B. analysed the data. M.Y., D.H., Y.L., T.C., C.S.W. and L.Z. synthesized lipid-DNA and other reagents. Y.L., M.Y., L.Q. and Q.L. prepared giant unilamellar vesicles and lipid monolayer. M.Y. and W.T. co-wrote the manuscript. W.T. supervised the project. All authors discussed the results and commented on the manuscript.

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Correspondence to Mingxu You or Weihong Tan.

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The authors declare no competing financial interests.

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You, M., Lyu, Y., Han, D. et al. DNA probes for monitoring dynamic and transient molecular encounters on live cell membranes. Nature Nanotech 12, 453–459 (2017). https://doi.org/10.1038/nnano.2017.23

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