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

文献类型： 外文期刊

作者： Zhang, Yaoyao 1 ; Luo, Jun 4 ; Tang, Na 1 ; Teng, Man 4 ; Reddy, Vishwanatha R. A. P. 1 ; Moffat, Katy 1 ; Shen, Zhiqi 1 ;

作者机构： 1.Pirbright Inst, Surrey GU24 0NF, England

2.UK China Ctr Excellence Res Avian Dis, Surrey GU24 0NF, England

3.Guangxi Univ, Coll Anim Sci & Technol, Nanning 530004, Peoples R China

4.Henan Acad Agr Sci, Henan Prov Key Lab Anim Immunol, Minist Agr, Key Lab Anim Immunol, Zhengzhou 450002, Henan, Peoples R China

5.Henan Univ Sci & Technol, Coll Anim Sci & Technol, Luoyang 471003, Peoples R China

6.Binzhou Anim Sci & Vet Med Acad, Binzhou 256600, Peoples R China

7.UK China Ctr Excellence Res Avian Dis, Binzhou 256600, Peoples R China

8.Univ Oxford, Jenner Inst, Old Rd Campus Res Bldg,Roosevelt Dr, Oxford OX3 7DQ, England

9.Univ Oxford, Dept Zool, 11a Mansfield Rd, Oxford OX1 3SZ, England

关键词： CRISPR; Cas9; MDV-transformed cell line; pp38; GFP; proliferation

期刊名称：VIRUSES-BASEL （ 影响因子：5.048； 五年影响因子：5.127 ）

ISSN： 1999-4915

年卷期： 2019 年 11 卷 5 期

页码：

收录情况： SCI

摘要： Marek's disease virus (MDV), a lymphotropic -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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