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Multi-Step Assembly of an RNA-Liposome Nanoparticle Formulation Revealed by Real-Time, Single-Particle Quantitative

Michael C Chung1,2, Hector R Mendez-Gomez3, Dhruvkumar Soni3

  • 1Division of Chemical Biology and Medicinal Chemistry, College of Pharmacy, University of Texas at Austin, Austin, TX, 78712, USA.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|January 31, 2025
PubMed
Summary

Researchers developed a new method to track nanoparticle self-assembly in real-time. This technique monitors liposome and mRNA interactions, crucial for mRNA cancer immunotherapy, revealing discrete assembly steps for improved nanomaterial design.

Keywords:
cancer‐therapydrug‐deliveryimagingnanomaterialsnanoparticlesquantitative imagingself‐assembly

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Area of Science:

  • Nanomaterial Science
  • Biotechnology
  • Immunotherapy

Background:

  • Self-assembly is vital for nanoparticle applications, but real-time, single-particle monitoring of multi-component systems remains difficult.
  • Current methods lack the throughput and model-independence needed for complex nanomaterial systems.

Purpose of the Study:

  • To develop and apply a novel multi-color fluorescence microscopy technique for real-time, high-throughput, single-particle tracking of multi-component nanomaterial self-assembly.
  • To investigate the discrete assembly steps of liposomes and messenger RNA (mRNA) in the context of mRNA-based cancer immunotherapy.

Main Methods:

  • Utilized multi-color fluorescence microscopy to simultaneously track liposome and mRNA assembly.
  • Analyzed assembly dynamics using a Smoluchowski model with a Brownian diffusion kernel.
  • Investigated the influence of RNA orientation on interaction transitions.

Main Results:

  • Observed two distinct assembly steps: initial RNA adsorption onto liposomes, followed by clustering into heterogeneous structures.
  • Characterized the size of assembled structures, ranging from sub-micrometer to tens of micrometers.
  • Demonstrated that the clustering process aligns with a Smoluchowski model and is influenced by RNA interaction orientation.

Conclusions:

  • The developed fluorescence microscopy approach provides a facile and effective method for monitoring multi-component nanomaterial self-assembly at the single-particle level.
  • This technique is broadly applicable to diverse nanomaterials beyond liposomes and mRNA, facilitating advancements in drug delivery and nanotechnology.
  • Understanding these assembly dynamics is critical for optimizing nanoparticle-based therapeutics, particularly in mRNA cancer immunotherapy.