MAP3K8 in Swine Testicular Cells Serves as an Anti-Viral Gene Against Porcine Delta Coronavirus

Abstract

Porcine deltacoronavirus (PDCoV) is the causative virus of an emerging porcine disease outbreak that has spread globally with up to 40% mortality in neonatal pigs. PDCoV has been shown to cross over and infect chickens and humans by attaching to the conserved regions in the host’s aminopeptidase N (APN) receptor. Therapeutic tools to combat a potential PDCoV outbreak can be developed by looking at genes commonly upregulated upon coronavirus infections. Mitogen-activated protein kinase kinase kinase 8 (MAP3K8) was a candidate gene upregulated in both PDCoV and SARS-CoV-2 infections in human cell lines. MAP3K8 is an intermediate enzyme involved in the MAP kinase pathway that activates from toll-like and interleukin 1 receptor activation, producing transcription factors for cytokine and growth factors. This study aims to investigate the role of MAP3K8 in the innate immunity of Swine Testicular (ST) cells in response to PDCoV infection. The CRISPR/Cas9 system was used to induce MAP3K8 knockout cell lines using a 3rd-generation lentivirus system to induce guide RNAs into the genome of ST cells with the Cas9 enzyme. Upon PDCoV infection, there was a significant increase in viral replication for the knockout MAP3K8 ST cell line, suggesting MAP3K8’s role as an antiviral factor. We hypothesize that this is due to MAP3K8 being involved in cytokine production, in which other immune cells react to and transcribe more proteins to induce apoptosis. This study presents a preliminary finding of a candidate antiviral protein that could be overexpressed to help mitigate PDCoV infection.

Advisor

Evans, Ellyn

Department

Biochemistry and Molecular Biology

Disciplines

Pathogenic Microbiology | Virology

Keywords

Coronavirus, Porcine Deltacoronavirus, sus scrofa, swine testicular cell, MAP3K8, pathogen-host interactions, CRISPR/Cas9, pathogenesis

Publication Date

2024

Degree Granted

Bachelor of Arts

Document Type

Senior Independent Study Thesis

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