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

Nature Nanotechnology volume 12, pages 453–459 (2017)Cite this article

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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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Author notes

  1. Mingxu You, Yifan Lyu and Da Han: These authors contributed equally to this work

Authors and Affiliations

  1. Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University, Changsha, 410082, People's Republic of China

    Mingxu You, Yifan Lyu, Da Han, Liping Qiu, Qiaoling Liu, Tao Chen, Cuichen Sam Wu, Liqin Zhang & Weihong Tan

  2. Department of Chemistry and Department of Physiology and Functional Genomics, Center for Research at the Bio/Nano Interface, Health Cancer Center, UF Genetics Institute, McKnight Brain Institute, University of Florida, Gainesville, 32611, Florida, USA

    Mingxu You, Yifan Lyu, Da Han, Liping Qiu, Tao Chen, Cuichen Sam Wu, Lu Peng, Liqin Zhang & Weihong Tan

  3. Department of Chemistry, University of Massachusetts Amherst, Amherst, 01003, Massachusetts, USA

    Mingxu You

  4. Department of Mathematics, Michigan State University, East Lansing, 48824, Michigan, USA

    Mingxu You

  5. School of Mathematical Sciences, Zhejiang University, Hangzhou, 310027, Zhejiang, People's Republic of China

    Gang Bao

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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.

Corresponding authors

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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