Latest Advances of Virology Research Using CRISPR/Cas9-Based Gene-Editing Technology and Its Application to Vaccine Development.

Man Teng,Yongxiu Yao,Jun Luo,Venugopal Nair

doi:10.3390/v13050779

Abstract

In recent years, the CRISPR/Cas9-based gene-editing techniques have been well developed and applied widely in several aspects of research in the biological sciences, in many species, including humans, animals, plants, and even in viruses. Modification of the viral genome is crucial for revealing gene function, virus pathogenesis, gene therapy, genetic engineering, and vaccine development. Herein, we have provided a brief review of the different technologies for the modification of the viral genomes. Particularly, we have focused on the recently developed CRISPR/Cas9-based gene-editing system, detailing its origin, functional principles, and touching on its latest achievements in virology research and applications in vaccine development, especially in large DNA viruses of humans and animals. Future prospects of CRISPR/Cas9-based gene-editing technology in virology research, including the potential shortcomings, are also discussed.

Highlights

In addition to the classical recombination and selection of recombinant viruses in virus-infected eukaryotic cells used previously [1], three main kinds of technology systems have been developed for viral genome engineering: (a) the bacteria-based homologous recombination system (BHRs) [2,3], (b) the bacterial artificial chromosome system (BACs) [4], and (c) the hybrid yeast–bacteria cloning system (HYCs) [5]
We have briefly reviewed the origination, functional principle of the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas system and focused on the latest advances of CRISPR/Cas9 system-based gene editing in virology and its applications in antiviral vaccine development, especially in large DNA viruses of animals
In 2015, some researchers cotransfected the purified pseudorabies virus (PRV) genomes with the constructed specific gRNA CRISPR/Cas9 complex into PK15 cells and obtained up to 100% viral gene editing efficiency [77]

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. In addition to the studies on gene function, gene therapy, and virus–host interaction, CRISPR/Cas9-based gene-editing technology has been widely applied in vaccine research for its high efficiency, specificity, versatility, flexibility, simplicity, and low cost compared to the other viral genome editing techniques, which has demonstrated an efficient and powerful strategy for future development of genetically engineered vaccines. Using the CRISPR/Cas9-based gene editing approach, a highly efficient recombinant virus, containing CDV virus-like particles (VLPs) and concurrently expressing the matrix (M), H, and F genes, has been recently constructed, which can assemble CDVVLPs and provide faster seroconversion and higher rates of antibody positivity than the parental virus strain among foxes and minks [120] This is the first report suggesting that the CRISPR/Cas system can be applied for rapid and efficient vaccine development for the prevention of CD among dogs, foxes, and minks in the future

Prospects

Findings

Methods