Genomics and Applied Biology 2024, Vol.15, No.5, 264-275 http://bioscipublisher.com/index.php/gab 264 Feature Review Open Access Harnessing Gene Editing Tools to Study ASFV Pathogenesis Xiaofang Lin Tropical Animal Medicine Center, Hainan Institute of Tropical Agricultural Resources, Sanya, 572024, Hainan, China Corresponding author: xiaofang.lin@hitar.org Genomics and Applied Biology, 2024, Vol.15, No.5 doi: 10.5376/gab.2024.15.0028 Received: 01 Sep., 2024 Accepted: 08 Oct., 2024 Published: 22 Oct., 2024 Copyright © 2024 Lin, This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Lin X.F., 2024, Harnessing gene editing tools to study ASFV pathogenesis, Genomics and Applied Biology, 15(5): 264-275 (doi: 10.5376/gab.2024.15.0028) Abstract The study utilizes advanced gene editing tools, specifically the CRISPR/Cas9 system, to investigate the pathogenesis of African Swine Fever Virus (ASFV) by creating recombinant virus strains with targeted gene deletions. The study successfully demonstrated the application of CRISPR/Cas9 to delete key immune response modulation genes (A238L, EP402R, and 9GL) in ASFV. The reconstituted virus exhibited similar replication kinetics to the parent virus, indicating that these genes can be modified with low frequency. Additionally, the use of CRISPR/Cas9 significantly accelerated the production of recombinant ASFV strains, reducing the time required from several months to less than two months. The study also highlighted the potential of CRISPR/Cas12a for sensitive and specific detection of ASFV, which could be crucial for on-site diagnostics and control of ASF outbreaks. The findings underscore the utility of CRISPR/Cas9 and CRISPR/Cas12a systems in both the study of ASFV pathogenesis and the development of rapid diagnostic tools. These advancements could pave the way for more effective control measures and the potential development of live-attenuated vaccines for ASFV. Keywords African Swine Fever Virus; CRISPR/Cas9; Gene editing; Pathogenesis; Diagnostic tools; Recombinant virus; ASFV detection 1 Introduction African Swine Fever Virus (ASFV) is a highly contagious and often lethal virus that causes hemorrhagic fever in domestic pigs and wild boar. The disease is characterized by high mortality rates, leading to severe economic consequences for the global swine industry. ASFV has been identified as one of the most significant threats to animal farming, with major outbreaks reported in Eastern Europe, Asia, and recently in India (Cackett et al., 2020; Senthilkumar et al., 2022). The virus's ability to spread rapidly and the lack of effective vaccines or antiviral treatments exacerbate its impact, making it a critical concern for animal health and food security worldwide (O'Donnell et al., 2016; Cackett et al., 2020). Controlling ASFV outbreaks is challenging due to several factors. Firstly, there are no commercially available vaccines or antiviral drugs to combat the virus, necessitating the culling of infected animals as the primary control measure (Cackett et al., 2020; Gladue et al., 2020). Secondly, the virus's complex genome and its ability to evade the host immune response complicate the development of effective vaccines (Gallardo et al., 2018; Bosch-Camós et al., 2021). Additionally, the virus's persistence in the environment and its ability to infect wild boar populations make eradication efforts difficult (Gallardo et al., 2023). These limitations highlight the urgent need for advanced research to understand the virus's biology and develop novel strategies for its control (O'Donnell et al., 2016; Cackett et al., 2020). Gene editing tools, such as CRISPR/Cas9, have revolutionized the field of virology by enabling precise modifications of viral genomes. These tools are particularly relevant for studying ASFV pathogenesis, as they allow researchers to investigate the functions of specific viral genes and their roles in virulence and immune evasion (O'Donnell et al., 2015; 2016). For instance, the deletion of certain ASFV genes has been shown to attenuate the virus, providing insights into potential vaccine candidates (O'Donnell et al., 2015; 2016; Gladue et al., 2020). By leveraging gene editing technologies, researchers can dissect the molecular mechanisms underlying ASFV infection and identify novel targets for therapeutic intervention (O'Donnell et al., 2016; Teklue et al., 2020).
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