Application of CRISPR/Cas9 in Understanding Avian Viruses and Developing Poultry Vaccines

Abstract

Clustered regularly interspaced short palindromic repeats associated protein nuclease 9 (CRISPR-Cas9) technology offers novel approaches to precisely, cost-effectively, and user-friendly edit genomes for a wide array of applications and across multiple disciplines. This methodology can be leveraged to underpin host-virus interactions, elucidate viral gene functions, and to develop recombinant vaccines. The successful utilization of CRISPR/Cas9 in editing viral genomes has paved the way of developing novel and multiplex viral vectored poultry vaccines. Furthermore, CRISPR/Cas9 can be exploited to rectify major limitations of conventional approaches including reversion to virulent form, recombination with field viruses and transgene, and genome instability. This review provides comprehensive analysis of the potential of CRISPR/Cas9 genome editing technique in understanding avian virus-host interactions and developing novel poultry vaccines. Finally, we discuss the simplest and practical aspects of genome editing approaches in generating multivalent recombinant poultry vaccines that conform simultaneous protection against major avian diseases.

Development of a Highly Efficient CRISPR/Cas9-Mediated Herpesvirus of Turkey-Based Vaccine against Novel Variant Infectious Bursal Disease Virus

“…More importantly, this approach could also be used to edit multiple genes at once [ 27 ]. Therefore, the use of this state-of-art technology could greatly accelerate multivalent vaccine development [ 13 , 21 ].…”

“…However, these technologies are often time consuming and labor-intensive. Recently, proof-of-concept studies have shown that the epoch-marking genome-engineering technology clustered regularly interspaced short palindromic repeats (CRISPR)/associated 9 (Cas9) is a rapid and efficient method for generating recombinants, which has spurred the development of effective poultry vaccines [ 11 , 12 , 13 , 14 ].…”

Methods for the Manipulation of Herpesvirus Genome and the Application to Marek’s Disease Virus Research

“…In a subsequent study, they inserted two additional viral antigen expression cassettes, which included genes of infectious laryngotracheitis virus (ILTV) and avian influenza virus (AIV), into the HVT-VP2 genome [121]. Theirs and other studies demonstrated that CRISPR/Cas9 is a simple and efficient method for the generation of vector vaccines carrying more than one foreign viral gene [122,123]. In addition, the nature of the CRISPR/Cas9 system promises the possibility of editing several target sites simultaneously, which will facilitate the development of recombinant vaccines that confer protection against multiple diseases.…”

CRISPR/Cas9 Editing of Duck Enteritis Virus Genome for the Construction of a Recombinant Vaccine Vector Expressing ompH Gene of Pasteurella multocida in Two Novel Insertion Sites

“…NHEJ is also faster than HDR, and it is also known to suppress the HDR process [ 34 ]. The combination of CRISPR/Cas9 and Cre–Lox recombination techniques provides a powerful and versatile system to excise pre-determined LoxP sites, allowing the excision of specific genetic fragments and selectable markers [ 26 , 35 ]. Through its simplicity and effectivity, CRISPR/Cas9 technology has shown to be beneficial for gene modification and provides an alternative to recombinant vaccine construction.…”