Staff View: A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice

A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice

Outbreaks of Pseudorabies (PR) by numerous highly virulent and antigenic variant Pseudorabies virus (PRV) strains have been causing severe economic losses to the pig industry in China since 2011. However, current commercial vaccines are often unable to induce thorough protective immunity. In this st...

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Main Authors:	Chen Luo, Qianqian Wang, Ruhai Guo, Jingnan Zhang, Jingya Zhang, Riteng Zhang, Xin Ma, Peixin Wang, Fathalrhman Eisa Addoma Adam, Basit Zeshan, Zengqi Yang, Yefei Zhou, Xinglong Wang
Format:	Article
Language:	English English
Published:	Elsevier B.V. 2022
Subjects:	RA638 Immunity and immunization in relation to public health SF1-1100 Animal culture
Online Access:	https://eprints.ums.edu.my/id/eprint/34792/1/Abstract.pdf https://eprints.ums.edu.my/id/eprint/34792/2/Full%20text.pdf https://eprints.ums.edu.my/id/eprint/34792/ https://www.sciencedirect.com/science/article/abs/pii/S0168170222002659 https://doi.org/10.1016/j.virusres.2022.198937
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spelling	my.ums.eprints.347922022-11-10T12:19:37Z https://eprints.ums.edu.my/id/eprint/34792/ A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice Chen Luo Qianqian Wang Ruhai Guo Jingnan Zhang Jingya Zhang Riteng Zhang Xin Ma Peixin Wang Fathalrhman Eisa Addoma Adam Basit Zeshan Zengqi Yang Yefei Zhou Xinglong Wang RA638 Immunity and immunization in relation to public health SF1-1100 Animal culture Outbreaks of Pseudorabies (PR) by numerous highly virulent and antigenic variant Pseudorabies virus (PRV) strains have been causing severe economic losses to the pig industry in China since 2011. However, current commercial vaccines are often unable to induce thorough protective immunity. In this study, a TK/gI/gE deleted recombinant PRV expressing GM-CSF was developed by using the HDR-CRISPR/Cas9 system. Here, a four-sgRNA along with the Cas9D10A targeting system was utilized for TK/gI/gE gene deletion and GM-CSF insertion. Our study showed that the four-sgRNA targeting system appeared to have higher knock-in efficiency for PRVs editing. The replication of the recombinant PRVs were slightly lower than that of the parental strain, but they appeared to have similar properties in terms of growth curves and plaque morphology. The mice vaccinated with the recombinant PRV expressing GM-CSF via intramuscular injection showed no obvious clinical symptoms, milder pathological lesions, and were completely protected against wild-type PRV challenge. When compared to the triple gene-deleted PRV, the gB antibodies and neutralizing antibody titers were improved and the immunized mice appeared to have lower viral load and higher mRNA levels of IL-2, IL-4, IL-6, and IFN-γ in spleens. Our study offers a novel approach for recombinant PRV construction, and the triple gene-deleted PRV expressing GM-CSF could serve as a promising vaccine candidate for PR control. Elsevier B.V. 2022 Article PeerReviewed text en https://eprints.ums.edu.my/id/eprint/34792/1/Abstract.pdf text en https://eprints.ums.edu.my/id/eprint/34792/2/Full%20text.pdf Chen Luo and Qianqian Wang and Ruhai Guo and Jingnan Zhang and Jingya Zhang and Riteng Zhang and Xin Ma and Peixin Wang and Fathalrhman Eisa Addoma Adam and Basit Zeshan and Zengqi Yang and Yefei Zhou and Xinglong Wang (2022) A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice. Virus Research, 322. pp. 1-35. ISSN 0168-1702 https://www.sciencedirect.com/science/article/abs/pii/S0168170222002659 https://doi.org/10.1016/j.virusres.2022.198937
institution	Universiti Malaysia Sabah
building	UMS Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaysia Sabah
content_source	UMS Institutional Repository
url_provider	http://eprints.ums.edu.my/
language	English English
topic	RA638 Immunity and immunization in relation to public health SF1-1100 Animal culture
spellingShingle	RA638 Immunity and immunization in relation to public health SF1-1100 Animal culture Chen Luo Qianqian Wang Ruhai Guo Jingnan Zhang Jingya Zhang Riteng Zhang Xin Ma Peixin Wang Fathalrhman Eisa Addoma Adam Basit Zeshan Zengqi Yang Yefei Zhou Xinglong Wang A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice
description	Outbreaks of Pseudorabies (PR) by numerous highly virulent and antigenic variant Pseudorabies virus (PRV) strains have been causing severe economic losses to the pig industry in China since 2011. However, current commercial vaccines are often unable to induce thorough protective immunity. In this study, a TK/gI/gE deleted recombinant PRV expressing GM-CSF was developed by using the HDR-CRISPR/Cas9 system. Here, a four-sgRNA along with the Cas9D10A targeting system was utilized for TK/gI/gE gene deletion and GM-CSF insertion. Our study showed that the four-sgRNA targeting system appeared to have higher knock-in efficiency for PRVs editing. The replication of the recombinant PRVs were slightly lower than that of the parental strain, but they appeared to have similar properties in terms of growth curves and plaque morphology. The mice vaccinated with the recombinant PRV expressing GM-CSF via intramuscular injection showed no obvious clinical symptoms, milder pathological lesions, and were completely protected against wild-type PRV challenge. When compared to the triple gene-deleted PRV, the gB antibodies and neutralizing antibody titers were improved and the immunized mice appeared to have lower viral load and higher mRNA levels of IL-2, IL-4, IL-6, and IFN-γ in spleens. Our study offers a novel approach for recombinant PRV construction, and the triple gene-deleted PRV expressing GM-CSF could serve as a promising vaccine candidate for PR control.
format	Article
author	Chen Luo Qianqian Wang Ruhai Guo Jingnan Zhang Jingya Zhang Riteng Zhang Xin Ma Peixin Wang Fathalrhman Eisa Addoma Adam Basit Zeshan Zengqi Yang Yefei Zhou Xinglong Wang
author_facet	Chen Luo Qianqian Wang Ruhai Guo Jingnan Zhang Jingya Zhang Riteng Zhang Xin Ma Peixin Wang Fathalrhman Eisa Addoma Adam Basit Zeshan Zengqi Yang Yefei Zhou Xinglong Wang
author_sort	Chen Luo
title	A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice
title_short	A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice
title_full	A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice
title_fullStr	A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice
title_full_unstemmed	A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice
title_sort	novel pseudorabies virus vaccine developed using hdr-crispr/cas9 induces strong humoral and cellular immune response in mice
publisher	Elsevier B.V.
publishDate	2022
url	https://eprints.ums.edu.my/id/eprint/34792/1/Abstract.pdf https://eprints.ums.edu.my/id/eprint/34792/2/Full%20text.pdf https://eprints.ums.edu.my/id/eprint/34792/ https://www.sciencedirect.com/science/article/abs/pii/S0168170222002659 https://doi.org/10.1016/j.virusres.2022.198937
_version_	1760231342494187520
score	13.211869

A novel Pseudorabies virus vaccine developed using HDR-CRISPR/Cas9 induces strong humoral and cellular immune response in mice

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