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Research Project: Countermeasures to Control and Eradicate Foreign Animal Diseases of Swine

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Title: CRISPR-Cas9, a tool to efficiently increase the development of recombinant African swine fever viruses

Author
item Borca, Manuel
item Holinka-Patterson, Lauren
item BERGGREN, KEITH - Oak Ridge Institute For Science And Education (ORISE)
item Gladue, Douglas

Submitted to: Scientific Reports
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/16/2018
Publication Date: 2/16/2018
Citation: Borca, M.V., Holinka-Patterson, L.G., Berggren, K., Gladue, D.P. 2018. CRISPR-Cas9, a tool to efficiently increase the development of recombinant African swine fever viruses. Scientific Reports. 8(3154). https://doi.org/10.1038/s41598-018-21575-8.
DOI: https://doi.org/10.1038/s41598-018-21575-8

Interpretive Summary: Making changes to viruses to the virus genome is an important aspect to study how viruses interact with the animals they infect. The difference between a live vaccine and a infectious virus often has to do with changes in the virus genome. In order to understand how a particular virus causes disease, and to make a rational vaccine, a researcher has to make genetic changes to the virus. For African swine fever virus making these changes has been very hard, and time consuming. However with the discovery of CRISPR-Cas9 a system to make genetic changes, this process has become easier in many instances. This paper describes the use of CRISPR-Cas9 in the field of African swine fever virus, and the ability to allow researchers make genetic changes in the viral genome, a dramatic improvement over traditional methods for genetically manipulating African swine fever virus.

Technical Abstract: African swine fever is a contagious and often lethal disease for domestic pigs with a significant economic impact on the swine industry. The etiological agent, African swine fever virus (ASFV), is a highly structurally complex double stranded DNA virus. No effective vaccines or antiviral treatment are currently commercially available. Development of recombinant ASFV for producing live-attenuated vaccines or studying the involvement of specific genes in virus virulence has relied on the relatively rare event of homologous recombination in primary swine macrophages, causing difficulty to purify the recombinant virus from the wild-type parental ASFV. Here we present the use of the CRISPR-Cas9 gene editing system as a more robust and efficient system to produce recombinant ASFVs.