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

Abstract Self‐assembly plays a critical role in nanoparticle‐based applications. However, it remains challenging to monitor the self‐assembly of multi‐component nanomaterials at a single‐particle level, in real‐time, with high throughput, and in a model‐independent manner. Here, multi‐color fluoresc...

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Main Authors: Michael C. Chung, Hector R. Mendez‐Gomez, Dhruvkumar Soni, Reagan McGinley, Alen Zacharia, Jewel Ashbrook, Brian Stover, Adam J. Grippin, Elias J. Sayour, Juan Guan
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Advanced Science
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Online Access:https://doi.org/10.1002/advs.202414305
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Summary:Abstract Self‐assembly plays a critical role in nanoparticle‐based applications. However, it remains challenging to monitor the self‐assembly of multi‐component nanomaterials at a single‐particle level, in real‐time, with high throughput, and in a model‐independent manner. Here, multi‐color fluorescence microscopy is applied to track the assembly of both liposomes and mRNA simultaneously in clinical mRNA‐based cancer immunotherapy. Imaging reveals that the assembly occurs in discrete steps: initially, RNA adsorbs onto the liposomes; then, the RNA‐coated liposomes cluster into heterogeneous structures ranging from sub‐micrometer to tens of micrometers. The clustering process is consistent with a Smoluchowski model with a Brownian diffusion kernel. The transition between the two steps of assembly is determined by the orientation of RNA‐mediated interactions. Given the facile application of this approach and the ubiquity of the components studied, the imaging and analysis in this work are readily applied to monitor multi‐component assembly of diverse nanomaterials.
ISSN:2198-3844