{"title":"CRISPR筛选揭示家蚕幼崽激素信号的全基因组调节因子。","authors":"Hao Sun, Chaojie Chen, Zijian Shi, Yue Liu, Ruolin Wang, Dan Liu, Tong Zhang, Sanyuan Ma, Ling Jia","doi":"10.1111/1744-7917.70103","DOIUrl":null,"url":null,"abstract":"<p><p>Insect metamorphosis constitutes a sophisticated endocrine cascade governed by the dynamic interplay between juvenile hormone (JH) and 20-hydroxyecdysone (20E). Although JH has been established as a critical developmental regulator, its intracellular signaling mechanisms remain incompletely characterized. In this study, we implemented a genome-wide CRISPR screening strategy in the lepidopteran model Bombyx mori to systematically identify components of JH signaling. First, we developed a JH-responsive reporter cell line with EGFP expression under the control of JH-sensitive promoters, enabling real-time monitoring of pathway activation. Then we established a genome-scale CRISPR knockout library specifically optimized for silkworm cells, and performed screening through integration of high-throughput flow cytometry and next-generation sequencing. Systematic analysis revealed a total of 387 candidate genes that significantly modulated JH signaling activity (231 enriched vs. 156 depleted). Functional annotation clustering revealed predominant enrichment in membrane biogenesis processes and small molecule metabolism. It is worth noting that through integrated genomic screening and experimental verification, we found that the tyrosine kinase receptor BmALK is an important regulatory node for JH signal transduction. This study establishes the first comprehensive genetic landscape of JH signaling in a holometabolous insect, providing a robust platform for functional mining of JH signaling pathways, and molecular evidence for membrane-associated JH signal transduction in Lepidoptera.</p>","PeriodicalId":13618,"journal":{"name":"Insect Science","volume":" ","pages":""},"PeriodicalIF":3.0000,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"CRISPR screening unravels genome-wide regulators of juvenile hormone signaling in Bombyx mori.\",\"authors\":\"Hao Sun, Chaojie Chen, Zijian Shi, Yue Liu, Ruolin Wang, Dan Liu, Tong Zhang, Sanyuan Ma, Ling Jia\",\"doi\":\"10.1111/1744-7917.70103\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Insect metamorphosis constitutes a sophisticated endocrine cascade governed by the dynamic interplay between juvenile hormone (JH) and 20-hydroxyecdysone (20E). Although JH has been established as a critical developmental regulator, its intracellular signaling mechanisms remain incompletely characterized. In this study, we implemented a genome-wide CRISPR screening strategy in the lepidopteran model Bombyx mori to systematically identify components of JH signaling. First, we developed a JH-responsive reporter cell line with EGFP expression under the control of JH-sensitive promoters, enabling real-time monitoring of pathway activation. Then we established a genome-scale CRISPR knockout library specifically optimized for silkworm cells, and performed screening through integration of high-throughput flow cytometry and next-generation sequencing. Systematic analysis revealed a total of 387 candidate genes that significantly modulated JH signaling activity (231 enriched vs. 156 depleted). Functional annotation clustering revealed predominant enrichment in membrane biogenesis processes and small molecule metabolism. It is worth noting that through integrated genomic screening and experimental verification, we found that the tyrosine kinase receptor BmALK is an important regulatory node for JH signal transduction. This study establishes the first comprehensive genetic landscape of JH signaling in a holometabolous insect, providing a robust platform for functional mining of JH signaling pathways, and molecular evidence for membrane-associated JH signal transduction in Lepidoptera.</p>\",\"PeriodicalId\":13618,\"journal\":{\"name\":\"Insect Science\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":3.0000,\"publicationDate\":\"2025-07-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Insect Science\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1111/1744-7917.70103\",\"RegionNum\":1,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ENTOMOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Insect Science","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1111/1744-7917.70103","RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ENTOMOLOGY","Score":null,"Total":0}
CRISPR screening unravels genome-wide regulators of juvenile hormone signaling in Bombyx mori.
Insect metamorphosis constitutes a sophisticated endocrine cascade governed by the dynamic interplay between juvenile hormone (JH) and 20-hydroxyecdysone (20E). Although JH has been established as a critical developmental regulator, its intracellular signaling mechanisms remain incompletely characterized. In this study, we implemented a genome-wide CRISPR screening strategy in the lepidopteran model Bombyx mori to systematically identify components of JH signaling. First, we developed a JH-responsive reporter cell line with EGFP expression under the control of JH-sensitive promoters, enabling real-time monitoring of pathway activation. Then we established a genome-scale CRISPR knockout library specifically optimized for silkworm cells, and performed screening through integration of high-throughput flow cytometry and next-generation sequencing. Systematic analysis revealed a total of 387 candidate genes that significantly modulated JH signaling activity (231 enriched vs. 156 depleted). Functional annotation clustering revealed predominant enrichment in membrane biogenesis processes and small molecule metabolism. It is worth noting that through integrated genomic screening and experimental verification, we found that the tyrosine kinase receptor BmALK is an important regulatory node for JH signal transduction. This study establishes the first comprehensive genetic landscape of JH signaling in a holometabolous insect, providing a robust platform for functional mining of JH signaling pathways, and molecular evidence for membrane-associated JH signal transduction in Lepidoptera.
期刊介绍:
Insect Science is an English-language journal, which publishes original research articles dealing with all fields of research in into insects and other terrestrial arthropods. Papers in any of the following fields will be considered: ecology, behavior, biogeography, physiology, biochemistry, sociobiology, phylogeny, pest management, and exotic incursions. The emphasis of the journal is on the adaptation and evolutionary biology of insects from the molecular to the ecosystem level. Reviews, mini reviews and letters to the editor, book reviews, and information about academic activities of the society are also published.