title: Depletion of elongation initiation factor 4E binding proteins by CRISPR/Cas9 enhances the antiviral response in porcine cells.
abstract: Type I interferons (IFNs) are key mediators of the innate antiviral response in mammalian cells. Elongation initiation factor 4E binding proteins (4E-BPs) are translational controllers of interferon regulatory factor 7 (IRF-7), the "master regulator" of IFN transcription. Previous studies have suggested that mouse cells depleted of 4E-BPs are more sensitive to IFNβ treatment and had lower viral loads as compared to wild type (WT) cells. However, such approach has not been tested as an antiviral strategy in livestock species. In this study, we tested the antiviral activity of porcine cells depleted of 4E-BP1 by a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) genome engineering system. We found that 4E-BP1 knockout (KO) porcine cells had increased expression of IFNα and β, IFN stimulated genes, and significant reduction in vesicular stomatitis virus titer as compare to WT cells. No phenotypical changes associated with CRISPR/Cas9 manipulation were observed in 4E-BP1 KO cells. This work highlights the use of the CRISPR/Cas9 system to enhance the antiviral response in porcine cells.
authors.affiliation: College of Veterinary Medicine and Biomedical Science, Texas A & M University, College Station, TX, USA; Plum Island Animal Disease Center (PIADC), Foreign Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Greenport, NY, USA; Oak Ridge Institute for Science and Education (ORISE), Plum Island Animal Disease Center (PIADC), Research Participation Program, Oak Ridge, TN, USA. Electronic address: Lisbeth.ramirez@ars.usda.gov.
authors.forename: Lisbeth
authors.initials: L
authors.lastname: Ramírez-Carvajal
authors.affiliation: College of Veterinary Medicine and Biomedical Science, Texas A & M University, College Station, TX, USA.
authors.forename: Neetu
authors.initials: N
authors.lastname: Singh
authors.affiliation: Plum Island Animal Disease Center (PIADC), Foreign Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Greenport, NY, USA.
authors.forename: Teresa
authors.initials: T
authors.lastname: de los Santos
authors.affiliation: Plum Island Animal Disease Center (PIADC), Foreign Animal Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, Greenport, NY, USA.
authors.forename: Luis L
authors.initials: LL
authors.lastname: Rodríguez
authors.affiliation: College of Veterinary Medicine and Biomedical Science, Texas A & M University, College Station, TX, USA.
authors.forename: Charles R
authors.initials: CR
authors.lastname: Long
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D064130:CRISPR-Associated Proteins / Q000378:metabolism*; D064113:CRISPR-Cas Systems; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D039561:Eukaryotic Initiation Factor-4E / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D055786:Gene Knockout Techniques; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics; D016899:Interferon-beta / Q000096:biosynthesis / Q000235:genetics; D013552:Swine; D014721:Vesicular stomatitis Indiana virus
chemical_list: D000998:Antiviral Agents; D064130:CRISPR-Associated Proteins; D039561:Eukaryotic Initiation Factor-4E; D016898:Interferon-alpha; D016899:Interferon-beta
keywords: 4E-BP1 knockout; CRISPR/Cas9; Interferon; Pig cells