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Related Concept Videos

MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
Nucleic Acid Structure01:25

Nucleic Acid Structure

The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms  a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA has a double-helix structure. The...

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Related Experiment Video

Updated: May 13, 2026

Generation of Cationic Nanoliposomes for the Efficient Delivery of In Vitro Transcribed Messenger RNA
08:29

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

Published on: February 1, 2019

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Transfection Potency of Lipid Nanoparticles Containing mRNA Depends on Relative Loading Levels.

Suiyang Liao1,2,3, Shuangyu Wang1, Abishek Wadhwa4,5

  • 1Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.

ACS Applied Materials & Interfaces
|December 31, 2024
PubMed
Summary
This summary is machine-generated.

Higher mRNA loading in lipid nanoparticles (LNPs) does not improve, but decreases, their ability to deliver genetic material. Fractionating LNPs by density revealed lower transfection efficiency in highly loaded particles.

Keywords:
encapsulationfractionationlipid nanoparticlesmRNAtherapeutics

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Area of Science:

  • Biotechnology
  • Nanomedicine
  • Molecular Biology

Background:

  • Lipid nanoparticles (LNPs) are crucial for mRNA delivery.
  • The impact of mRNA loading levels on LNP function remains largely unknown.
  • Understanding LNP formulation is key to optimizing mRNA therapeutics.

Purpose of the Study:

  • To investigate the relationship between mRNA loading levels and LNP properties.
  • To determine if mRNA loading affects LNP functionality in vitro and in vivo.
  • To explore LNP fractionation based on mRNA content.

Main Methods:

  • Formulation of mRNA into lipid nanoparticles (LNPs) with varying mRNA copy numbers.
  • Fractionation of LNPs using ultracentrifugation based on distinct mass densities.
  • Characterization of LNP size, lipid composition, and morphology post-fractionation.
  • In vitro and in vivo assessment of mRNA delivery and transfection efficiency.

Main Results:

  • LNPs exhibited distinct mass densities correlating with mRNA loading levels.
  • Fractionation via ultracentrifugation separated LNPs based on mRNA content.
  • No significant correlation was found between mRNA loading and LNP size, lipid composition, or morphology.
  • The LNP fraction with the highest mRNA loading demonstrated the lowest transfection competence.

Conclusions:

  • mRNA loading level is a critical factor influencing LNP functional delivery.
  • Highly mRNA-loaded LNPs are less effective at delivering their genetic payload.
  • Fractionation strategies may be necessary to isolate optimal LNP populations for mRNA therapeutics.