Cargando…

Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori

The silkworm Bombyx mori is an economically important insect. The sericulture industry is seriously affected by pathogen infections. Of these pathogens, Bombyx mori nucleopolyhedrovirus (BmNPV) causes approximately 80% of the total economic losses due to pathogen infections. We previously constructe...

Descripción completa

Detalles Bibliográficos
Autores principales: Liu, Yujia, Chen, Dongbin, Zhang, Xiaoqian, Chen, Shuqing, Yang, Dehong, Tang, Linmeng, Yang, Xu, Wang, Yaohui, Luo, Xingyu, Wang, Manli, Hu, Zhihong, Huang, Yongping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8780094/
https://www.ncbi.nlm.nih.gov/pubmed/35062262
http://dx.doi.org/10.3390/v14010059
_version_ 1784637746923438080
author Liu, Yujia
Chen, Dongbin
Zhang, Xiaoqian
Chen, Shuqing
Yang, Dehong
Tang, Linmeng
Yang, Xu
Wang, Yaohui
Luo, Xingyu
Wang, Manli
Hu, Zhihong
Huang, Yongping
author_facet Liu, Yujia
Chen, Dongbin
Zhang, Xiaoqian
Chen, Shuqing
Yang, Dehong
Tang, Linmeng
Yang, Xu
Wang, Yaohui
Luo, Xingyu
Wang, Manli
Hu, Zhihong
Huang, Yongping
author_sort Liu, Yujia
collection PubMed
description The silkworm Bombyx mori is an economically important insect. The sericulture industry is seriously affected by pathogen infections. Of these pathogens, Bombyx mori nucleopolyhedrovirus (BmNPV) causes approximately 80% of the total economic losses due to pathogen infections. We previously constructed a BmNPV-specific CRISPR/Cas9 silkworm line with significantly enhanced resistance to BmNPV. In order to optimize the resistance properties and minimize its impact on economic traits, we constructed an inducible CRISPR/Cas9 system for use in transgenic silkworms. We used the 39k promoter, which is induced by viral infection, to express Cas9 and the U6 promoter to express four small guide RNA targeting the genes encoding BmNPV late expression factors 1 and 3 (lef-1 and lef-3, respectively), which are essential for viral DNA replication. The system was rapidly activated when the silkworm was infected and showed considerably higher resistance to BmNPV infection than the wild-type silkworm. The inducible system significantly reduced the development effects due to the constitutive expression of Cas9. No obvious differences in developmental processes or economically important characteristics were observed between the resulting transgenic silkworms and wild-type silkworms. Adoption of this accurate and highly efficient inducible CRISPR/Cas9 system targeting BmNPV DNA replication will result in enhanced antivirus measures during sericulture, and our work also provides insights into the broader application of the CRISPR/Cas9 system in the control of infectious diseases and insect pests.
format Online
Article
Text
id pubmed-8780094
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-87800942022-01-22 Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori Liu, Yujia Chen, Dongbin Zhang, Xiaoqian Chen, Shuqing Yang, Dehong Tang, Linmeng Yang, Xu Wang, Yaohui Luo, Xingyu Wang, Manli Hu, Zhihong Huang, Yongping Viruses Article The silkworm Bombyx mori is an economically important insect. The sericulture industry is seriously affected by pathogen infections. Of these pathogens, Bombyx mori nucleopolyhedrovirus (BmNPV) causes approximately 80% of the total economic losses due to pathogen infections. We previously constructed a BmNPV-specific CRISPR/Cas9 silkworm line with significantly enhanced resistance to BmNPV. In order to optimize the resistance properties and minimize its impact on economic traits, we constructed an inducible CRISPR/Cas9 system for use in transgenic silkworms. We used the 39k promoter, which is induced by viral infection, to express Cas9 and the U6 promoter to express four small guide RNA targeting the genes encoding BmNPV late expression factors 1 and 3 (lef-1 and lef-3, respectively), which are essential for viral DNA replication. The system was rapidly activated when the silkworm was infected and showed considerably higher resistance to BmNPV infection than the wild-type silkworm. The inducible system significantly reduced the development effects due to the constitutive expression of Cas9. No obvious differences in developmental processes or economically important characteristics were observed between the resulting transgenic silkworms and wild-type silkworms. Adoption of this accurate and highly efficient inducible CRISPR/Cas9 system targeting BmNPV DNA replication will result in enhanced antivirus measures during sericulture, and our work also provides insights into the broader application of the CRISPR/Cas9 system in the control of infectious diseases and insect pests. MDPI 2021-12-30 /pmc/articles/PMC8780094/ /pubmed/35062262 http://dx.doi.org/10.3390/v14010059 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Liu, Yujia
Chen, Dongbin
Zhang, Xiaoqian
Chen, Shuqing
Yang, Dehong
Tang, Linmeng
Yang, Xu
Wang, Yaohui
Luo, Xingyu
Wang, Manli
Hu, Zhihong
Huang, Yongping
Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori
title Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori
title_full Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori
title_fullStr Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori
title_full_unstemmed Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori
title_short Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori
title_sort construction of baculovirus-inducible crispr/cas9 antiviral system targeting bmnpv in bombyx mori
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8780094/
https://www.ncbi.nlm.nih.gov/pubmed/35062262
http://dx.doi.org/10.3390/v14010059
work_keys_str_mv AT liuyujia constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT chendongbin constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT zhangxiaoqian constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT chenshuqing constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT yangdehong constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT tanglinmeng constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT yangxu constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT wangyaohui constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT luoxingyu constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT wangmanli constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT huzhihong constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori
AT huangyongping constructionofbaculovirusinduciblecrisprcas9antiviralsystemtargetingbmnpvinbombyxmori