DNA-Mediated Liposome Fusion Observed by Fluorescence Spectrometry

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

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

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.

Original languageEnglish
Title of host publicationNucleic Acid Detection and Structural Investigations : Methods and Protocols
EditorsKira Astakhova, Syeda Atia Bukhari
Number of pages18
Volume2063
PublisherHumana Press
Publication date2020
Pages101-118
Chapter9
ISBN (Print)978-1-0716-0137-2
ISBN (Electronic)978-1-0716-0138-9
DOIs
Publication statusPublished - 2020
SeriesMethods in Molecular Biology
Volume2063
ISSN1064-3745

Keywords

  • Controlled fusion
  • Fluorometry
  • Lipidated DNA
  • Liposome

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