Single-particle combinatorial multiplexed liposome fusion mediated by DNA

Nat Chem. 2022 May;14(5):558-565. doi: 10.1038/s41557-022-00912-5. Epub 2022 Apr 4.

Abstract

Combinatorial high-throughput methodologies are central for both screening and discovery in synthetic biochemistry and biomedical sciences. They are, however, often reliant on large-scale analyses and thus limited by a long running time and excessive materials cost. We here present a single-particle combinatorial multiplexed liposome fusion mediated by DNA for parallelized multistep and non-deterministic fusion of individual subattolitre nanocontainers. We observed directly the efficient (>93%) and leakage free stochastic fusion sequences for arrays of surface-tethered target liposomes with six freely diffusing populations of cargo liposomes, each functionalized with individual lipidated single-stranded DNA and fluorescently barcoded by a distinct ratio of chromophores. The stochastic fusion resulted in a distinct permutation of fusion sequences for each autonomous nanocontainer. Real-time total internal reflection imaging allowed the direct observation of >16,000 fusions and 566 distinct fusion sequences accurately classified using machine learning. The high-density arrays of surface-tethered target nanocontainers (~42,000 containers per mm2) offers entire combinatorial multiplex screens using only picograms of material.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA*
  • DNA, Single-Stranded
  • Liposomes*
  • Membrane Fusion

Substances

  • DNA, Single-Stranded
  • Liposomes
  • DNA