Facile Synthesis of N-Doped Graphene Quantum Dots as Novel Transfection Agents for mRNA and pDNA.

Minchul Ahn,Jaekwang Song,Byung Hee Hong

doi:10.3390/nano11112816

Abstract

In the wake of the coronavirus disease 2019 (COVID-19) pandemic, global pharmaceutical companies have developed vaccines for the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Some have adopted lipid nanoparticles (LNPs) or viral vectors to deliver the genes associated with the spike protein of SARS-CoV-2 for vaccination. This strategy of vaccination by delivering genes to express viral proteins has been successfully applied to the mRNA vaccines for COVID-19, and is also applicable to gene therapy. However, conventional transfection agents such as LNPs and viral vectors are not yet sufficient to satisfy the levels of safety, stability, and efficiency required for the clinical applications of gene therapy. In this study, we synthesized N-doped graphene quantum dots (NGQDs) for the transfection of various genes, including messenger ribonucleic acids (mRNAs) and plasmid deoxyribonucleic acids (pDNAs). The positively charged NGQDs successfully formed electrostatic complexes with negatively charged mRNAs and pDNAs, and resulted in the efficient delivery and transfection of the genes into target cells. The transfection efficiency of NGQDs is found to be comparable to that of commercially available LNPs. Considering their outstanding stability even at room temperature as well as their low toxicity, NGQDs are expected to be novel universal gene delivery platforms that can outperform LNPs and viral vectors.

Highlights

COVID-19, coronavirus disease 2019, has threatened global society, and various types of vaccines have been developed to overcome the pandemic [1,2,3,4,5,6]
From the overall characterization data, we demonstrated that N-doped graphene quantum dots (NGQDs) had the characteristic properties of graphene quantum dots (GQDs): the graphitic core with the diverse functional groups and the positive charge required to interact with genes
green fluorescent protein (GFP) in the NGQDs + messenger ribonucleic acids (mRNAs) complex group was comparable to groups using Lipofecratio, and the results show that 30 ng of mRNA, encoding a GFP, formed complexes with 600 ng of NGQDs before transfection

Summary

Introduction

COVID-19, coronavirus disease 2019, has threatened global society, and various types of vaccines have been developed to overcome the pandemic [1,2,3,4,5,6]. The therapeutic strategy that uses genes such as mRNA or plasmid DNA (pDNA) is called gene therapy, which has drawn much attention because it expresses desired proteins in the body through their corresponding sequence information to be transcribed and translated into therapeutic proteins to fight various diseases [16,17,18]. The viral vectors such as adeno-associated virus (AAV) are the main platforms that deliver genes into cells with high transfection efficiency [19,20].

Methods

Discussion

Conclusion