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

1
The Pirbright Institute & UK-China Centre of Excellence for Research on Avian Diseases, Pirbright, Surrey GU24 0NF, UK
2
College of Animal Science and Technology, Guangxi University, Nanning 530004, China
3
Key Laboratory of Animal Immunology of the Ministry of Agriculture, Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
4
College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China
5
Binzhou Animal Science and Veterinary Medicine Academy & UK-China Centre of Excellence for Research on Avian Diseases, Binzhou 256600, China
6
The Jenner Institute, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK
7
Department of Zoology, University of Oxford, 11a Mansfield Road, Oxford OX1 3SZ, UK
*
Authors to whom correspondence should be addressed.
Viruses 2019, 11(5), 391; https://doi.org/10.3390/v11050391
Received: 27 March 2019 / Revised: 23 April 2019 / Accepted: 24 April 2019 / Published: 26 April 2019
(This article belongs to the Section Animal Viruses)
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Abstract

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. View Full-Text
Keywords: CRISPR/Cas9; MDV-transformed cell line; pp38; GFP; proliferation CRISPR/Cas9; MDV-transformed cell line; pp38; GFP; proliferation
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Zhang, Y.; Luo, J.; Tang, N.; Teng, M.; Reddy, V.R.; Moffat, K.; Shen, Z.; Nair, V.; Yao, Y. Targeted Editing of the pp38 Gene in Marek’s Disease Virus-Transformed Cell Lines Using CRISPR/Cas9 System. Viruses 2019, 11, 391.

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