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Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector

Turkey herpesvirus (HVT) has been widely used as a successful live virus vaccine against Marek's disease (MD) in chickens for more than five decades. Increasingly, HVT is also used as a highly effective recombinant vaccine vector against multiple avian pathogens. Conventional recombination, or...

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Autores principales: Zai, Xusheng, Shi, Bin, Shao, Hongxia, Qian, Kun, Ye, Jianqiang, Yao, Yongxiu, Nair, Venugopal, Qin, Aijian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9174942/
https://www.ncbi.nlm.nih.gov/pubmed/35694305
http://dx.doi.org/10.3389/fmicb.2022.886873
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author Zai, Xusheng
Shi, Bin
Shao, Hongxia
Qian, Kun
Ye, Jianqiang
Yao, Yongxiu
Nair, Venugopal
Qin, Aijian
author_facet Zai, Xusheng
Shi, Bin
Shao, Hongxia
Qian, Kun
Ye, Jianqiang
Yao, Yongxiu
Nair, Venugopal
Qin, Aijian
author_sort Zai, Xusheng
collection PubMed
description Turkey herpesvirus (HVT) has been widely used as a successful live virus vaccine against Marek's disease (MD) in chickens for more than five decades. Increasingly, HVT is also used as a highly effective recombinant vaccine vector against multiple avian pathogens. Conventional recombination, or recombineering, techniques that involve the cloning of viral genomes and, more recently, gene editing methods have been used for the generation of recombinant HVT-based vaccines. In this study, we used NHEJ-dependent CRISPR/Cas9-based approaches to insert the mCherry cassette for the screening of the HVT genome and identifying new potential sites for the insertion of foreign genes. A novel intergenic site HVT-005/006 in the unique long (UL) region of the HVT genome was identified, and mCherry was found to be stably expressed when inserted at this site. To confirm whether this site was suitable for the insertion of other exogenous genes, haemagglutinin (HA) of the H9N2 virus was inserted into this site, and a recombinant HVT-005/006-HA was rescued. The recombinant HVT-HA can grow well and express HA protein stably, which demonstrated that HVT-005/006 is a promising site for the insertion of foreign genes.
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spelling pubmed-91749422022-06-09 Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector Zai, Xusheng Shi, Bin Shao, Hongxia Qian, Kun Ye, Jianqiang Yao, Yongxiu Nair, Venugopal Qin, Aijian Front Microbiol Microbiology Turkey herpesvirus (HVT) has been widely used as a successful live virus vaccine against Marek's disease (MD) in chickens for more than five decades. Increasingly, HVT is also used as a highly effective recombinant vaccine vector against multiple avian pathogens. Conventional recombination, or recombineering, techniques that involve the cloning of viral genomes and, more recently, gene editing methods have been used for the generation of recombinant HVT-based vaccines. In this study, we used NHEJ-dependent CRISPR/Cas9-based approaches to insert the mCherry cassette for the screening of the HVT genome and identifying new potential sites for the insertion of foreign genes. A novel intergenic site HVT-005/006 in the unique long (UL) region of the HVT genome was identified, and mCherry was found to be stably expressed when inserted at this site. To confirm whether this site was suitable for the insertion of other exogenous genes, haemagglutinin (HA) of the H9N2 virus was inserted into this site, and a recombinant HVT-005/006-HA was rescued. The recombinant HVT-HA can grow well and express HA protein stably, which demonstrated that HVT-005/006 is a promising site for the insertion of foreign genes. Frontiers Media S.A. 2022-05-25 /pmc/articles/PMC9174942/ /pubmed/35694305 http://dx.doi.org/10.3389/fmicb.2022.886873 Text en Copyright © 2022 Zai, Shi, Shao, Qian, Ye, Yao, Nair and Qin. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Zai, Xusheng
Shi, Bin
Shao, Hongxia
Qian, Kun
Ye, Jianqiang
Yao, Yongxiu
Nair, Venugopal
Qin, Aijian
Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector
title Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector
title_full Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector
title_fullStr Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector
title_full_unstemmed Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector
title_short Identification of a Novel Insertion Site HVT-005/006 for the Generation of Recombinant Turkey Herpesvirus Vector
title_sort identification of a novel insertion site hvt-005/006 for the generation of recombinant turkey herpesvirus vector
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9174942/
https://www.ncbi.nlm.nih.gov/pubmed/35694305
http://dx.doi.org/10.3389/fmicb.2022.886873
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