The Ins and Outs of Messenger RNA Electroporation for Physical Gene Delivery in Immune Cell-Based Therapy.

Diana Campillo-Davo,Donovan Flumens,Zwi N Berneman,Maxime De Laere,Viggo F I Van Tendeloo,Maarten Versteven,Eva Lion,Gils Roex,Sébastien Anguille

doi:10.3390/pharmaceutics13030396

Diana Campillo-Davo, Donovan Flumens + Show 7 more

Open Access

https://doi.org/10.3390/pharmaceutics13030396

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Journal: Pharmaceutics	Publication Date: Mar 16, 2021
Citations: 20	License type: CC BY 4.0

Affiliation: University of Antwerp, Antwerp University Hospital

Abstract

Messenger RNA (mRNA) electroporation is a powerful tool for transient genetic modification of cells. This non-viral method of genetic engineering has been widely used in immunotherapy. Electroporation allows fine-tuning of transfection protocols for each cell type as well as introduction of multiple protein-coding mRNAs at once. As a pioneering group in mRNA electroporation, in this review, we provide an expert overview of the ins and outs of mRNA electroporation, discussing the different parameters involved in mRNA electroporation as well as the production of research-grade and production and application of clinical-grade mRNA for gene transfer in the context of cell-based immunotherapies.

Highlights

Electroporation of messenger RNA (mRNA) is a versatile methodology for the transient expression of proteins of interest
As a highly flexible system, it allows the fine-tuning of transfection conditions for each cell type and to multiplex mRNAs as required
There is a wide variety of options when it comes to improving both the electroporation conditions and stability/translation of the mRNAs for monocistronic and polycistronic gene transfer

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. The field of cell-based immunotherapy in particular has made enormous progress due to the development and optimization of messenger RNA (mRNA) electroporation for gene transfer. This type of genetic engineering, compared to that of the viral delivery of genes, represents a safer alternative for protein expression with no risk of insertional mutagenesis and lower immunogenicity [4,5]. Several factors may influence the transfection efficiency (blue), synthesis (red) and translation (green) of mRNA in electroporation-based therapies These factors can be individually optimized, combined and tailored for each type of immune cell and target gene to be transferred.

The Physics

The Chemistry

The Biology

Clinical Production of mRNA for Electroporation

Clinical Application of mRNA Electroporation in Cell-Based Immunotherapies

Conclusions