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Related Experiment Video

Updated: Jun 9, 2025

Characterizing Extracellular Vesicles from Biological Fluids

Characterizing Extracellular Vesicles from Biological Fluids

Published on: February 28, 2025

Cationized extracellular vesicles for gene delivery.

Natalia L Klyachko^1,2, Matthew J Haney^3,4, Anton V Lopukhov⁵

¹Division of Pharmacoengineering and Molecular Pharmaceutics, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599-7362, USA. nlklyachko@gmail.com.

Scientific Reports

|October 29, 2024

Summary

Engineered extracellular vesicles (EVs) with cationic lipids efficiently deliver nucleic acids like plasmid DNA, mRNA, and siRNA into cancer cells and macrophages, enabling gene delivery applications.

Keywords:

Cancer EVs Gene delivery Multivalent cationic lipid Transfection

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

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

Published on: February 1, 2019

Area of Science:

Biotechnology
Nanomedicine
Molecular Biology

Background:

Extracellular vesicles (EVs) are recognized as effective drug delivery vehicles.
Previous research established EV-based systems for proteins and chemotherapeutics.
Nucleic acid delivery remains a challenge in therapeutic applications.

Purpose of the Study:

To engineer extracellular vesicles (EVs) for efficient nucleic acid delivery.
To investigate the use of multivalent cationic lipids in EV engineering.
To evaluate the delivery and transfection capabilities of engineered EVs for DNA, mRNA, and siRNA.

Main Methods:

Engineering EVs with multivalent cationic lipids.
Loading engineered EVs with plasmid DNA (pDNA), mRNA, or siRNA.
Assessing cellular uptake and intracellular delivery via confocal microscopy.
Evaluating transfection efficiency in vitro for pDNA and mRNA.
Measuring gene knockdown efficacy using siRNA in reporter gene assays.

Main Results:

Stable, small-sized cationized EVs were successfully produced.
Engineered EVs demonstrated efficient uptake by triple negative breast cancer (TNBC) cells and macrophages.
Delivered nucleic acid cargo reached the nuclei of target cells.
Significant in vitro transfection was achieved with pDNA- and mRNA-loaded EVs.
siRNA-loaded EVs effectively knocked down reporter gene expression.

Conclusions:

Multivalent cationized EVs are a novel and effective platform for nucleic acid delivery.
Engineered EVs show promise for gene therapy and gene silencing applications.
This strategy advances the potential of EVs in genetic medicine.