Genomics and Applied Biology 2024, Vol.15, No.5, 264-275 http://bioscipublisher.com/index.php/gab 269 5.2 Identification of potential drug targets Gene editing has been instrumental in pinpointing specific ASFV proteins and pathways that can serve as potential drug targets. For example, the identification of ASFV proteins such as P34, E199L, MGF360-15R, and E248R, which are involved in critical steps of the viral life cycle, has opened new avenues for antiviral drug development. These proteins interact with host cellular machinery, including Rab proteins that regulate the endocytic pathway, making them attractive targets for therapeutic intervention (García-Dorival et al., 2023). Furthermore, the discovery of ASFV genes that manipulate the interferon response, such as A276R, A528R, and I329L, highlights their potential as targets for antiviral drugs aimed at enhancing the host immune response (Correia et al., 2013). These findings underscore the power of gene editing in identifying and validating novel drug targets for ASFV. 5.3 Implications for vaccine development Gene editing technologies hold significant promise for informing the design and development of effective ASFV vaccines. By enabling the precise deletion or modification of specific viral genes, researchers can create attenuated virus strains that elicit a robust immune response without causing disease. For instance, the deletion of ASFV genes involved in immune evasion, such as those that inhibit the interferon response, could lead to the development of attenuated virus strains that are more immunogenic and capable of inducing protective immunity (Correia et al., 2013). Additionally, the identification of key antigenic regions on ASFV structural proteins, such as p54, through epitope mapping studies, provides valuable information for the design of subunit vaccines that target these critical regions (Petrovan et al., 2020). These advancements highlight the potential of gene editing to revolutionize ASFV vaccine development and contribute to the control and eradication of this devastating disease. 6 Case Studies and Current Research 6.1 Case study 1: CRISPR/Cas9 and ASFV resistance in pigs Recent advancements in gene editing technologies, particularly the CRISPR/Cas9 system, have shown promise in developing resistance to African swine fever virus (ASFV) in pigs. One notable study attempted to create a recombinant ASFV strain by deleting specific genes associated with the virus's ability to evade the host immune response. The genes A238L, EP402R, and 9GL were targeted using the CRISPR/Cas9 system. The modified virus demonstrated similar replication kinetics to the parent virus, indicating the potential of CRISPR/Cas9 in developing ASFV-resistant strains (Woźniakowski et al., 2020). Another study successfully generated a vaccine prototype by deleting the EP402R (CD2v) and A238L genes from the ASFV Arm/07/CBM/c2 strain using CRISPR/Cas9, which proved to be safe and fully protective against a virulent Korean Paju strain (Pérez-Núñez et al., 2022). 6.2 Case study 2: gene editing to study ASFV-host interactions Gene editing tools have also been instrumental in elucidating the interactions between ASFV and its host. For instance, the deletion of ASFV genes DP148R, DP71L, and DP96R from the highly virulent ASFV CN/GS/2018 strain resulted in significant attenuation of the virus. This mutant strain not only provided complete protection against homologous challenges in pigs but also revealed important insights into host-virus interactions. RNA sequencing analysis showed that the deletion of these genes led to the upregulation of the host histone H3.1 gene and downregulation of the ASFV MGF110-7L gene, highlighting potential targets for future therapeutic strategies (Qi et al., 2023). 6.3 Case study 3: high-throughput screening of ASFV genes High-throughput screening methods have been employed to identify critical ASFV genes that could serve as targets for vaccine development. One study developed a new live-attenuated vaccine candidate by deleting the H240R and MGF505-7R genes from the ASFV HLJ/18 strain. This mutant strain exhibited decreased viral titers in porcine alveolar macrophages and provided 100% protection in piglets against a virulent ASFV challenge, demonstrating the effectiveness of high-throughput gene screening in identifying potential vaccine candidates (Li et al., 2023) (Figure 3). Additionally, the use of CRISPR-Cas12a coupled with nucleic acid amplification has been optimized for the sensitive detection of ASFV, showcasing the potential of high-throughput screening in diagnostic applications (Tao et al., 2020).
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