Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance

Bovine viral diarrhea virus’s (BVDV) entry into bovine cells involves attachment of virions to cellular receptors, internalization, and pH-dependent fusion with endosomal membranes. The primary host receptor for BVDV is CD46; however, the complete set of host factors required for virus entry is unkn...

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Autores principales:	Aspen M. Workman, Michael P. Heaton, Dennis A. Webster, Gregory P. Harhay, Theodore S. Kalbfleisch, Timothy P. L. Smith, Shollie M. Falkenberg, Daniel F. Carlson, Tad S. Sonstegard
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	BVDV CRISPR PTPN12 GRID2 RABGAP1L CRIB Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/6a76f48d5448417cb8f382903301c78d
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spelling	oai:doaj.org-article:6a76f48d5448417cb8f382903301c78d2021-11-25T19:12:45ZEvaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance10.3390/v131121471999-4915https://doaj.org/article/6a76f48d5448417cb8f382903301c78d2021-10-01T00:00:00Zhttps://www.mdpi.com/1999-4915/13/11/2147https://doaj.org/toc/1999-4915Bovine viral diarrhea virus’s (BVDV) entry into bovine cells involves attachment of virions to cellular receptors, internalization, and pH-dependent fusion with endosomal membranes. The primary host receptor for BVDV is CD46; however, the complete set of host factors required for virus entry is unknown. The Madin-Darby bovine kidney (MDBK) cell line is susceptible to BVDV infection, while a derivative cell line (CRIB) is resistant at the level of virus entry. We performed complete genome sequencing of each to identify genomic variation underlying the resistant phenotype with the aim of identifying host factors essential for BVDV entry. Three large compound deletions in the BVDV-resistant CRIB cell line were identified and predicted to disrupt the function or expression of the genes <i>PTPN12</i>, <i>GRID2</i>, and <i>RABGAP1L</i>. However, CRISPR/Cas9 mediated knockout of these genes, individually or in combination, in the parental MDBK cell line did not impact virus entry or replication. Therefore, resistance to BVDV in the CRIB cell line is not due to the apparent spontaneous loss of <i>PTPN12</i>, <i>GRID2</i>, or <i>RABGAP1L</i> gene function. Identifying the functional cause of BVDV resistance in the CRIB cell line may require more detailed comparisons of the genomes and epigenomes.Aspen M. WorkmanMichael P. HeatonDennis A. WebsterGregory P. HarhayTheodore S. KalbfleischTimothy P. L. SmithShollie M. FalkenbergDaniel F. CarlsonTad S. SonstegardMDPI AGarticleBVDVCRISPRPTPN12GRID2RABGAP1LCRIBMicrobiologyQR1-502ENViruses, Vol 13, Iss 2147, p 2147 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	BVDV CRISPR PTPN12 GRID2 RABGAP1L CRIB Microbiology QR1-502
spellingShingle	BVDV CRISPR PTPN12 GRID2 RABGAP1L CRIB Microbiology QR1-502 Aspen M. Workman Michael P. Heaton Dennis A. Webster Gregory P. Harhay Theodore S. Kalbfleisch Timothy P. L. Smith Shollie M. Falkenberg Daniel F. Carlson Tad S. Sonstegard Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance
description	Bovine viral diarrhea virus’s (BVDV) entry into bovine cells involves attachment of virions to cellular receptors, internalization, and pH-dependent fusion with endosomal membranes. The primary host receptor for BVDV is CD46; however, the complete set of host factors required for virus entry is unknown. The Madin-Darby bovine kidney (MDBK) cell line is susceptible to BVDV infection, while a derivative cell line (CRIB) is resistant at the level of virus entry. We performed complete genome sequencing of each to identify genomic variation underlying the resistant phenotype with the aim of identifying host factors essential for BVDV entry. Three large compound deletions in the BVDV-resistant CRIB cell line were identified and predicted to disrupt the function or expression of the genes <i>PTPN12</i>, <i>GRID2</i>, and <i>RABGAP1L</i>. However, CRISPR/Cas9 mediated knockout of these genes, individually or in combination, in the parental MDBK cell line did not impact virus entry or replication. Therefore, resistance to BVDV in the CRIB cell line is not due to the apparent spontaneous loss of <i>PTPN12</i>, <i>GRID2</i>, or <i>RABGAP1L</i> gene function. Identifying the functional cause of BVDV resistance in the CRIB cell line may require more detailed comparisons of the genomes and epigenomes.
format	article
author	Aspen M. Workman Michael P. Heaton Dennis A. Webster Gregory P. Harhay Theodore S. Kalbfleisch Timothy P. L. Smith Shollie M. Falkenberg Daniel F. Carlson Tad S. Sonstegard
author_facet	Aspen M. Workman Michael P. Heaton Dennis A. Webster Gregory P. Harhay Theodore S. Kalbfleisch Timothy P. L. Smith Shollie M. Falkenberg Daniel F. Carlson Tad S. Sonstegard
author_sort	Aspen M. Workman
title	Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance
title_short	Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance
title_full	Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance
title_fullStr	Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance
title_full_unstemmed	Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance
title_sort	evaluating large spontaneous deletions in a bovine cell line selected for bovine viral diarrhea virus resistance
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/6a76f48d5448417cb8f382903301c78d
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Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance

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