Extracellular Nanovesicle Enhanced Gene Transfection Using Polyethyleneimine in HEK293T Cells and Zebrafish Embryos.

Zhenzhen Zhang,Chao Zhang,Fabrice Laroche,Zunfeng Liu,Xiang Zhou,Kai Wen,Zhenglong Wang,Jan Pieter Abrahams,Qiang Zhou

doi:10.3389/fbioe.2020.00448

Abstract

It is a hot topic to improve efficiency and decrease toxicity of gene transfection reagents. The extracellular nanovesicles (EVs) that are released by cells play an important role in intercellular communication and are naturally designed for genetic exchange between cells. Here, we show that the EVs have a large beneficial effect in polyethyleneimine (PEI)-mediated transfection of a GFP-encoding plasmid into HEK293T cells. An improvement of transfection efficiency of ~500% and a decrease in toxicity were observed in a specific concentration range of PEI. The EVs also greatly improved the transfection of the same plasmid into zebrafish embryos. To verify the generality of this gene transfection approach, we also tested the cell viability and gene transfection efficiency using two other plasmids (EpTEN and ELuc) and in another cell line (A549). The measured increase in transfection efficiency makes EV a promising candidate for enhancement of the quality of current PEI-based transfection technique.

Highlights

The importance of gene transfection has grown immensely over the past years, as gene therapy may remedy many diseases caused by genetic disorders
We investigated the effect of extracellular nanovesicles (EVs) for enhancing the gene transfection of PEI in mammalian cells and zebrafish embryos
This process was performed multiple times so that enough EVs were collected for gene transfection

Summary

Introduction

The importance of gene transfection has grown immensely over the past years, as gene therapy may remedy many diseases caused by genetic disorders. Gene transfection vectors can help genes to overcome cellular barriers, which include synthetic and viral vectors (Saffari et al, 2016). Viral vectors and their clinical trials in human gene therapy have saved human lives (Poletti et al, 2018). Viral vectors show high transfection efficiency, while they exhibit low gene-carrying capacity and limited cell-targeting specify (Hernandez-Garcia et al, 2014). The public health implications of each viral vector remain to be estimated on a case-by-case basis (Alessia et al, 2013)

Results

Discussion

Conclusion