CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive Pest
<i>Spodoptera frugiperda</i> (J. E. Smith), an emerging invasive pest worldwide, has posed a serious agricultural threat to the newly invaded areas. Although somatic sex differentiation is fundamentally conserved among insects, the sex determination cascade in <i>S. frugiperda</...
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MDPI AG
2022-11-01
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| author | Junwen Gu Jingyi Wang Honglun Bi Xuehai Li Austin Merchant Porui Zhang Qi Zhang Xuguo Zhou |
| author_facet | Junwen Gu Jingyi Wang Honglun Bi Xuehai Li Austin Merchant Porui Zhang Qi Zhang Xuguo Zhou |
| author_sort | Junwen Gu |
| collection | DOAJ |
| description | <i>Spodoptera frugiperda</i> (J. E. Smith), an emerging invasive pest worldwide, has posed a serious agricultural threat to the newly invaded areas. Although somatic sex differentiation is fundamentally conserved among insects, the sex determination cascade in <i>S. frugiperda</i> is largely unknown. In this study, we cloned and functionally characterized <i>Doublesex</i> (<i>dsx</i>), a “molecular switch” modulating sexual dimorphism in <i>S. frugiperda</i> using male- and female-specific isoforms. Given that Lepidoptera is recalcitrant to RNAi, CRISPR/Cas9-mediated mutagenesis was employed to construct <i>S. frugiperda</i> mutants. Specifically, we designed target sites on exons 2, 4, and 5 to eliminate the common, female-specific, and male-specific regions of <i>S. frugiperda dsx</i> (<i>Sfdsx</i>), respectively. As expected, abnormal development of both the external and internal genitalia was observed during the pupal and adult stages. Interestingly, knocking out sex-specific <i>dsx</i> variants in <i>S. frugiperda</i> led to significantly reduced fecundity and fertility in adults of corresponding sex. Our combined results not only confirm the conserved function of <i>dsx</i> in <i>S. frugiperda</i> sex differentiation but also provide empirical evidence for <i>dsx</i> as a potential target for the Sterile Insect Technique (SIT) to combat this globally invasive pest in a sustainable and environmentally friendly way. |
| first_indexed | 2024-03-09T18:25:46Z |
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| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T18:25:46Z |
| publishDate | 2022-11-01 |
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| spelling | doaj.art-cd0d7883cfcf4205bc8c79f30234dd642023-11-24T07:57:22ZengMDPI AGCells2073-44092022-11-011122355710.3390/cells11223557CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive PestJunwen Gu0Jingyi Wang1Honglun Bi2Xuehai Li3Austin Merchant4Porui Zhang5Qi Zhang6Xuguo Zhou7College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Plant Protection, Shenyang Agricultural University, Shenyang 110866, ChinaState Key Laboratory of Cotton Biology, School of Life Sciences, College of Agriculture, Henan University, Kaifeng 475004, ChinaCollege of Plant Protection, Shenyang Agricultural University, Shenyang 110866, ChinaDepartment of Entomology, University of Kentucky, Lexington, KY 40546, USACollege of Plant Protection, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Plant Protection, Shenyang Agricultural University, Shenyang 110866, ChinaDepartment of Entomology, University of Kentucky, Lexington, KY 40546, USA<i>Spodoptera frugiperda</i> (J. E. Smith), an emerging invasive pest worldwide, has posed a serious agricultural threat to the newly invaded areas. Although somatic sex differentiation is fundamentally conserved among insects, the sex determination cascade in <i>S. frugiperda</i> is largely unknown. In this study, we cloned and functionally characterized <i>Doublesex</i> (<i>dsx</i>), a “molecular switch” modulating sexual dimorphism in <i>S. frugiperda</i> using male- and female-specific isoforms. Given that Lepidoptera is recalcitrant to RNAi, CRISPR/Cas9-mediated mutagenesis was employed to construct <i>S. frugiperda</i> mutants. Specifically, we designed target sites on exons 2, 4, and 5 to eliminate the common, female-specific, and male-specific regions of <i>S. frugiperda dsx</i> (<i>Sfdsx</i>), respectively. As expected, abnormal development of both the external and internal genitalia was observed during the pupal and adult stages. Interestingly, knocking out sex-specific <i>dsx</i> variants in <i>S. frugiperda</i> led to significantly reduced fecundity and fertility in adults of corresponding sex. Our combined results not only confirm the conserved function of <i>dsx</i> in <i>S. frugiperda</i> sex differentiation but also provide empirical evidence for <i>dsx</i> as a potential target for the Sterile Insect Technique (SIT) to combat this globally invasive pest in a sustainable and environmentally friendly way.https://www.mdpi.com/2073-4409/11/22/3557<i>Spodoptera frugiperda</i>invasive species<i>doublesex</i>sex determinationCRISPR/Cas9 systemSIT |
| spellingShingle | Junwen Gu Jingyi Wang Honglun Bi Xuehai Li Austin Merchant Porui Zhang Qi Zhang Xuguo Zhou CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive Pest Cells <i>Spodoptera frugiperda</i> invasive species <i>doublesex</i> sex determination CRISPR/Cas9 system SIT |
| title | CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive Pest |
| title_full | CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive Pest |
| title_fullStr | CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive Pest |
| title_full_unstemmed | CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive Pest |
| title_short | CRISPR/Cas9-Mediated Mutagenesis of Sex-Specific <i>Doublesex</i> Splicing Variants Leads to Sterility in <i>Spodoptera frugiperda</i>, a Global Invasive Pest |
| title_sort | crispr cas9 mediated mutagenesis of sex specific i doublesex i splicing variants leads to sterility in i spodoptera frugiperda i a global invasive pest |
| topic | <i>Spodoptera frugiperda</i> invasive species <i>doublesex</i> sex determination CRISPR/Cas9 system SIT |
| url | https://www.mdpi.com/2073-4409/11/22/3557 |
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