Mechanistic insights into mRNA-LNP interactions: role of ionizable lipid content in regulating mRNA intracellular

Related Experiment Video

Updated: Apr 13, 2026

Author Spotlight: Enhancing Lipid Nanoparticle Formation Through Turbulent Mixing in Confined Geometries
08:10

Author Spotlight: Enhancing Lipid Nanoparticle Formation Through Turbulent Mixing in Confined Geometries

Published on: August 23, 2024

6.8K

Hoang Quan Truong1, Urmila Kafle1,2, Cao Thuy Giang Nguyen1

  • 1Department of Biomedical and Nutritional Sciences, University of Massachusetts Lowell, 3 Solomont Way, Lowell, 01854, MA, USA.

Journal of Nanobiotechnology
|April 11, 2026
PubMed

Insights

Optimizing lipid nanoparticles (LNPs) by reducing ionizable lipids enhances messenger RNA (mRNA) delivery and expression. Lower ionizable lipid content improves mRNA release from LNPs, boosting therapeutic potential.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Drug Delivery

Background:

  • Lipid nanoparticles (LNPs) are crucial for delivering messenger RNA (mRNA) therapeutics.
  • Intracellular mRNA release from LNPs is a significant, yet understudied, hurdle for effective translation.

Purpose of the Study:

  • To investigate the impact of ionizable lipid content in LNP formulations on mRNA delivery and translation.
  • To identify optimal ionizable lipid ratios for enhanced LNP-mediated mRNA performance.

Main Methods:

  • Systematic variation of ionizable lipid (SM-102, ALC-0315) molar ratios in LNP formulations.
  • Assessment of in vitro transfection efficiency and in vivo mRNA expression.
  • Analysis of cellular uptake, endosomal escape, and intracellular mRNA release via fluorescence colocalization.
  • Evaluation of antigen-specific immune responses in mice.

Main Results:

  • LNPs with lower ionizable lipid content (~30 mol%) showed significantly higher in vitro transfection and in vivo expression compared to standard formulations (50 mol% or 46.3 mol%).
  • Enhanced performance was observed despite comparable cellular uptake and endosomal escape.
  • Reduced ionizable lipid levels facilitated greater cytosolic mRNA release, indicating improved intracellular dissociation.
  • Optimized LNPs induced stronger humoral and cellular immune responses in mice.

Conclusions:

  • Intracellular mRNA-LNP dissociation is a critical determinant of LNP-mediated mRNA delivery efficiency, potentially more so than endosomal escape.
  • Fine-tuning the ionizable lipid composition of LNPs can substantially improve mRNA delivery, expression, and immunogenicity.
  • These findings offer a strategy for developing more effective mRNA therapeutics.
Keywords:
DissociationEndosomal escapeIntracellular releaseLNPsmRNAmRNA-LNP interaction

More Related Videos

Testing the In Vitro and In Vivo Efficiency of mRNA-Lipid Nanoparticles Formulated by Microfluidic Mixing08:55

Testing the In Vitro and In Vivo Efficiency of mRNA-Lipid Nanoparticles Formulated by Microfluidic Mixing

Published on: January 20, 2023

15.2K
Generation of Cationic Nanoliposomes for the Efficient Delivery of In Vitro Transcribed Messenger RNA08:29

Generation of Cationic Nanoliposomes for the Efficient Delivery of In Vitro Transcribed Messenger RNA

Published on: February 1, 2019

10.8K

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...
7.3K
Regulated mRNA Transport02:22

Regulated mRNA Transport

3.5K
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
1.6K
Translational Regulation01:29

Translational Regulation

Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
859
Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
27.4K
Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
3.5K