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Related Experiment Video

Updated: Jul 7, 2025

Bead Loading Proteins and Nucleic Acids into Adherent Human Cells

Bead Loading Proteins and Nucleic Acids into Adherent Human Cells

Published on: June 1, 2021

A Method for Using Cell-Penetrating Peptides for Loading Plasmid DNA into Secreted Extracellular Vesicles.

Jekaterina Nebogatova¹, Heleri Heike Härk¹, Anett Puskar¹

¹Institute of Technology, University of Tartu, Nooruse 1, 50411 Tartu, Estonia.

|December 23, 2023

Summary

This summary is machine-generated.

This study introduces a novel method for loading plasmid DNA (pDNA) into extracellular vesicles (EVs) using cell-penetrating peptides (CPPs). This approach significantly enhances pDNA delivery and expression in recipient cells, reducing toxicity for in vivo applications.

Keywords:

cell-penetrating peptides drug delivery system extracellular vesicles gene therapy nucleic acid therapeutics

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Last Updated: Jul 7, 2025

Bead Loading Proteins and Nucleic Acids into Adherent Human Cells

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Peptide-derived Method to Transport Genes and Proteins Across Cellular and Organellar Barriers in Plants

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Engineering Cell-permeable Protein

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Published on: December 28, 2009

Area of Science:

Biotechnology
Molecular Biology
Nanomedicine

Background:

Plasmid DNA (pDNA) therapeutics face challenges due to low bioavailability and high toxicity.
Extracellular vesicles (EVs) are promising biocompatible carriers for nucleic acid delivery.
Existing methods for loading pDNA into EVs have shown limited success.

Purpose of the Study:

To develop a novel and efficient method for loading pDNA into EVs.
To evaluate the transfection efficacy and bioavailability of pDNA delivered via EVs.
To assess the in vivo toxicity of pDNA-loaded EVs compared to traditional delivery methods.

Main Methods:

Loading EVs with pDNA via transient transfection using cell-penetrating peptides (CPPs).
Quantifying reporter gene expression (luciferase) in recipient cells.
Evaluating in vivo toxicity and therapeutic effects of pDNA-loaded EVs.

Main Results:

A 10^4-fold increase in luciferase expression was observed in recipient cells.
Demonstrated high transfection efficacy and bioavailability of encapsulated nucleic acids.
In vivo studies showed reduced toxicity of pDNA-loaded EVs compared to traditional nucleic acid delivery.

Conclusions:

Transient transfection with CPPs provides an effective method for pDNA loading into EVs.
pDNA-loaded EVs offer enhanced bioavailability and transfection efficiency for in vivo applications.
EVs serve as safe and effective native delivery vehicles, mitigating the toxicity of pDNA therapeutics.