Insights into the mechanisms of electromediated gene delivery and application to the loading of giant vesicles with negatively charged macromolecules

TL;DR

This study investigates how electric fields facilitate DNA entry into giant vesicles, demonstrating electrophoresis as the main mechanism and proposing a predictive model for efficient encapsulation of negatively charged molecules.

Contribution

The paper introduces a quantitative model for DNA electrotransfer into vesicles, highlighting electrophoresis as the key mechanism and enabling high-efficiency loading of charged macromolecules.

Findings

Electrotransfer of DNA into GUVs is primarily via electrophoresis.

A simple theoretical model accurately predicts DNA transfer based on electric field parameters.

The method enables efficient encapsulation of negatively charged macromolecules.

Abstract

We present experimental results regarding the electrotransfer of plasmid DNA into phosphatidylcholine giant unilamellar vesicles (GUVs). Our observations indicate that a direct entry is the predominant mechanism of electrotransfer. A quantitative analysis of the DNA concentration increments inside the GUVs is also performed, and we find that our experimental data are very well described by a simple theoretical model in which DNA entry is mostly driven by electrophoresis. Our theoretical framework allows for the prediction of the amount of transfered DNA as a function of the electric field parameters, and thus paves the way towards a novel method for encapsulating with high efficiency not only DNA, but any negatively charged macromolecule into GUVs.