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Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System

Marek’s disease virus (MDV), a lymphotropic α-herpesvirus associated with T-cell lymphomas in chickens, is an excellent model for herpesvirus biology and virus-induced oncogenesis. Marek’s disease (MD) is also one of the cancers against which a vaccine was first used. In the lymphomas and lymphoblas...

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Autores principales: Zhang, Yaoyao, Luo, Jun, Tang, Na, Teng, Man, Reddy, Vishwanatha R.A.P., Moffat, Katy, Shen, Zhiqiang, Nair, Venugopal, Yao, Yongxiu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6563304/
https://www.ncbi.nlm.nih.gov/pubmed/31027375
http://dx.doi.org/10.3390/v11050391
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author Zhang, Yaoyao
Luo, Jun
Tang, Na
Teng, Man
Reddy, Vishwanatha R.A.P.
Moffat, Katy
Shen, Zhiqiang
Nair, Venugopal
Yao, Yongxiu
author_facet Zhang, Yaoyao
Luo, Jun
Tang, Na
Teng, Man
Reddy, Vishwanatha R.A.P.
Moffat, Katy
Shen, Zhiqiang
Nair, Venugopal
Yao, Yongxiu
author_sort Zhang, Yaoyao
collection PubMed
description Marek’s disease virus (MDV), a lymphotropic α-herpesvirus associated with T-cell lymphomas in chickens, is an excellent model for herpesvirus biology and virus-induced oncogenesis. Marek’s disease (MD) is also one of the cancers against which a vaccine was first used. In the lymphomas and lymphoblastoid cell lines (LCLs) derived from them, MDV establishes latent infection with limited gene expression. Although LCLs are valuable for interrogating viral and host gene functions, molecular determinants associated with the maintenance of MDV latency and lytic switch remain largely unknown, mainly due to the lack of tools for in situ manipulation of the genomes in these cell lines. Here we describe the first application of CRISPR/Cas9 editing approach for precise editing of the viral gene phosphoprotein 38 (pp38), a biomarker for latent/lytic switch in MDV-transformed LCLs MDCC-MSB-1 (Marek’s disease cell line MSB-1) and MDCC-HP8. Contradictory to the previous reports suggesting that pp38 is involved in the maintenance of transformation of LCL MSB-1 cells, we show that pp38-deleted cells proliferated at a significant higher rate, suggesting that pp38 is dispensable for the transformed state of these cell lines. Application of CRISPR/Cas9-based gene editing of MDV-transformed cell lines in situ opens up further opportunities towards a better understanding of MDV pathogenesis and virus-host interactions.
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spelling pubmed-65633042019-06-17 Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System Zhang, Yaoyao Luo, Jun Tang, Na Teng, Man Reddy, Vishwanatha R.A.P. Moffat, Katy Shen, Zhiqiang Nair, Venugopal Yao, Yongxiu Viruses Article Marek’s disease virus (MDV), a lymphotropic α-herpesvirus associated with T-cell lymphomas in chickens, is an excellent model for herpesvirus biology and virus-induced oncogenesis. Marek’s disease (MD) is also one of the cancers against which a vaccine was first used. In the lymphomas and lymphoblastoid cell lines (LCLs) derived from them, MDV establishes latent infection with limited gene expression. Although LCLs are valuable for interrogating viral and host gene functions, molecular determinants associated with the maintenance of MDV latency and lytic switch remain largely unknown, mainly due to the lack of tools for in situ manipulation of the genomes in these cell lines. Here we describe the first application of CRISPR/Cas9 editing approach for precise editing of the viral gene phosphoprotein 38 (pp38), a biomarker for latent/lytic switch in MDV-transformed LCLs MDCC-MSB-1 (Marek’s disease cell line MSB-1) and MDCC-HP8. Contradictory to the previous reports suggesting that pp38 is involved in the maintenance of transformation of LCL MSB-1 cells, we show that pp38-deleted cells proliferated at a significant higher rate, suggesting that pp38 is dispensable for the transformed state of these cell lines. Application of CRISPR/Cas9-based gene editing of MDV-transformed cell lines in situ opens up further opportunities towards a better understanding of MDV pathogenesis and virus-host interactions. MDPI 2019-04-26 /pmc/articles/PMC6563304/ /pubmed/31027375 http://dx.doi.org/10.3390/v11050391 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhang, Yaoyao
Luo, Jun
Tang, Na
Teng, Man
Reddy, Vishwanatha R.A.P.
Moffat, Katy
Shen, Zhiqiang
Nair, Venugopal
Yao, Yongxiu
Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System
title Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System
title_full Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System
title_fullStr Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System
title_full_unstemmed Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System
title_short Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System
title_sort targeted editing of the pp38 gene in marek’s disease virus-transformed cell lines using crispr/cas9 system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6563304/
https://www.ncbi.nlm.nih.gov/pubmed/31027375
http://dx.doi.org/10.3390/v11050391
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