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DNA-Mediated Liposome Fusion Observed by Fluorescence Spectrometry

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Nucleic Acid Detection and Structural Investigations

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2063))

Abstract

DNA-programmed and controlled fusion of lipid membranes have recently been optimized to reliably mix the contents between two populations of liposomes, each functionalized with complementary lipidated DNA (LiNA) oligomer. In this chapter we describe a procedure for DNA-controlled fusion of liposomes mediated by LiNAs that are designed to force bilayers into close proximity. Using a self-quenching fluorescent dye (Sulforhodamine B) to monitor both the mixing of the internal volumes and leakage of the dye into the outer volume we measure the efficiency of content mixing in the bulk population, allowing for direct comparison between different LiNA designs. By generating samples for calibration corresponding to different amounts of content mixing, the average number of fusion events per labeled liposome can be estimated.

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Acknowledgments

We thank Dr. Alexander Rabe for his contributions to method development and verification. The authors gratefully acknowledge funding by the Biomolecular Nanoscale Engineering Center (BioNEC), a Centre of Excellence funded by The VILLUM Foundation, grant no. VKR022710.

Conflicts of interest: The authors have no conflicts of interest to declare.

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Authors and Affiliations

  1. Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense M, Denmark

    Philipp M. G. Löffler, Oliver Ries & Stefan Vogel

Authors
  1. Philipp M. G. Löffler
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  2. Oliver Ries
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  3. Stefan Vogel
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Correspondence to Stefan Vogel .

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Editors and Affiliations

  1. Department of Chemistry, Technical University of Denmark, Kongens Lyngby, Denmark

    Kira Astakhova

  2. Department of Chemistry, Technical University of Denmark, Kongens Lyngby, Denmark

    Syeda Atia Bukhari

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Löffler, P.M.G., Ries, O., Vogel, S. (2020). DNA-Mediated Liposome Fusion Observed by Fluorescence Spectrometry. In: Astakhova, K., Bukhari, S. (eds) Nucleic Acid Detection and Structural Investigations. Methods in Molecular Biology, vol 2063. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0138-9_9

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  • DOI: https://doi.org/10.1007/978-1-0716-0138-9_9

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