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Related Concept Videos

Regulated mRNA Transport02:22

Regulated mRNA Transport

In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...
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A Quantitative Fluorescence Microscopy-based Single Liposome Assay for Detecting the Compositional Inhomogeneity Between Individual Liposomes
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Single-Particle Multiparametric Microscopy Reveals Structural, Size, and Payload Heterogeneity in mRNA-Loaded Lipid

Albert Kamanzi1,2,3, Ariadne Tuckmantel Bido1,3, Yao Zhang1,4,5

  • 1Michael Smith Laboratories, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.

ACS Nano
|December 19, 2025
PubMed
Summary

Researchers developed a new microscopy technique to analyze mRNA-containing lipid nanoparticles (LNPs). This method reveals detailed LNP properties, crucial for designing effective nanomedicines and vaccines.

Keywords:
FRETdrug deliverylipid nanoparticlesmRNA payloadnanomedicinessingle-molecule microscopy

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Area of Science:

  • Nanomedicine
  • Biophysics
  • Microscopy

Background:

  • Understanding lipid nanoparticle (LNP) heterogeneity is key for therapeutic applications.
  • Microscopic properties of LNPs correlate with their biological function.

Purpose of the Study:

  • To develop and apply a novel microscopy technique for simultaneous measurement of LNP properties.
  • To investigate the relationship between LNP formulation and microscopic characteristics.

Main Methods:

  • Combined alternating laser excitation (ALEX) with convex lens-induced confinement (CLiC) microscopy.
  • Simultaneously measured size, multicolor fluorescence, mRNA payload, and Förster resonance energy transfer (FRET) of individual LNPs.
  • Varied formulation parameters including ionizable lipids, buffers, and molecular ratios.

Main Results:

  • Per-particle lipid fluorescence was lower for empty vs. mRNA-loaded LNPs.
  • The relative size of empty vs. mRNA-loaded LNPs depended on formulation and internal structure.
  • CLiC-ALEX revealed heterogeneity in mRNA copy number and structural arrangements within LNPs.

Conclusions:

  • The developed CLiC-ALEX technique provides rigorous biophysical insights into LNP heterogeneity.
  • These findings are critical for understanding structure-activity relationships in nanomedicines.
  • The study informs the rational design of advanced nanomedicines and vaccine formulations.