CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in Silkworms
<i>Bombyx mori</i> nucleopolyhedrovirus (BmNPV) is a pathogen that causes severe disease in silkworms. In a previous study, we demonstrated that by using the CRISPR/Cas9 system to disrupt the BmNPV <i>ie-1</i> and <i>me53</i> genes, transgenic silkworms showed res...
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MDPI AG
2022-05-01
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| author | Yujia Liu Xiaoqian Zhang Dongbin Chen Dehong Yang Chenxu Zhu Linmeng Tang Xu Yang Yaohui Wang Xingyu Luo Manli Wang Yongping Huang Zhihong Hu Zulian Liu |
| author_facet | Yujia Liu Xiaoqian Zhang Dongbin Chen Dehong Yang Chenxu Zhu Linmeng Tang Xu Yang Yaohui Wang Xingyu Luo Manli Wang Yongping Huang Zhihong Hu Zulian Liu |
| author_sort | Yujia Liu |
| collection | DOAJ |
| description | <i>Bombyx mori</i> nucleopolyhedrovirus (BmNPV) is a pathogen that causes severe disease in silkworms. In a previous study, we demonstrated that by using the CRISPR/Cas9 system to disrupt the BmNPV <i>ie-1</i> and <i>me53</i> genes, transgenic silkworms showed resistance to BmNPV infection. Here, we used the same strategy to simultaneously target <i>lef8</i> and <i>lef9</i>, which are essential for BmNPV replication. A PCR assay confirmed that double-stranded breaks were induced in viral DNA at targeted sequences in BmNPV-infected transgenic silkworms that expressed small guide RNAs (sgRNAs) and Cas9. Bioassays and qPCR showed that replication of BmNPV and mortality were significantly reduced in the transgenic silkworms in comparison with the control groups. Microscopy showed degradation of midgut cells in the BmNPV-infected wild type silkworms, but not in the transgenic silkworms. These results demonstrated that transgenic silkworms using the CRISPR/Cas9 system to disrupt BmNPV <i>lef8</i> and <i>lef9</i> genes could successfully prevent BmNPV infection. Our research not only provides more alternative targets for the CRISPR antiviral system, but also aims to provide new ideas for the application of virus infection research and the control of insect pests. |
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| id | doaj.art-aa46fd77157f44fdb4be1cc613d1c526 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T22:15:34Z |
| publishDate | 2022-05-01 |
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| record_format | Article |
| series | Viruses |
| spelling | doaj.art-aa46fd77157f44fdb4be1cc613d1c5262023-11-23T19:23:54ZengMDPI AGViruses1999-49152022-05-01146111910.3390/v14061119CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in SilkwormsYujia Liu0Xiaoqian Zhang1Dongbin Chen2Dehong Yang3Chenxu Zhu4Linmeng Tang5Xu Yang6Yaohui Wang7Xingyu Luo8Manli Wang9Yongping Huang10Zhihong Hu11Zulian Liu12Key Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaState Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, ChinaState Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, ChinaKey Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China<i>Bombyx mori</i> nucleopolyhedrovirus (BmNPV) is a pathogen that causes severe disease in silkworms. In a previous study, we demonstrated that by using the CRISPR/Cas9 system to disrupt the BmNPV <i>ie-1</i> and <i>me53</i> genes, transgenic silkworms showed resistance to BmNPV infection. Here, we used the same strategy to simultaneously target <i>lef8</i> and <i>lef9</i>, which are essential for BmNPV replication. A PCR assay confirmed that double-stranded breaks were induced in viral DNA at targeted sequences in BmNPV-infected transgenic silkworms that expressed small guide RNAs (sgRNAs) and Cas9. Bioassays and qPCR showed that replication of BmNPV and mortality were significantly reduced in the transgenic silkworms in comparison with the control groups. Microscopy showed degradation of midgut cells in the BmNPV-infected wild type silkworms, but not in the transgenic silkworms. These results demonstrated that transgenic silkworms using the CRISPR/Cas9 system to disrupt BmNPV <i>lef8</i> and <i>lef9</i> genes could successfully prevent BmNPV infection. Our research not only provides more alternative targets for the CRISPR antiviral system, but also aims to provide new ideas for the application of virus infection research and the control of insect pests.https://www.mdpi.com/1999-4915/14/6/1119BmNPVCRISPR/Cas9transgenic silkwormantiviral therapy |
| spellingShingle | Yujia Liu Xiaoqian Zhang Dongbin Chen Dehong Yang Chenxu Zhu Linmeng Tang Xu Yang Yaohui Wang Xingyu Luo Manli Wang Yongping Huang Zhihong Hu Zulian Liu CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in Silkworms Viruses BmNPV CRISPR/Cas9 transgenic silkworm antiviral therapy |
| title | CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in Silkworms |
| title_full | CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in Silkworms |
| title_fullStr | CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in Silkworms |
| title_full_unstemmed | CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in Silkworms |
| title_short | CRISPR/Cas9-Mediated Disruption of the <i>lef8</i> and <i>lef9</i> to Inhibit Nucleopolyhedrovirus Replication in Silkworms |
| title_sort | crispr cas9 mediated disruption of the i lef8 i and i lef9 i to inhibit nucleopolyhedrovirus replication in silkworms |
| topic | BmNPV CRISPR/Cas9 transgenic silkworm antiviral therapy |
| url | https://www.mdpi.com/1999-4915/14/6/1119 |
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