CRISPR筛选揭示家蚕幼崽激素信号的全基因组调节因子。

IF 3 1区 农林科学 Q1 ENTOMOLOGY
Hao Sun, Chaojie Chen, Zijian Shi, Yue Liu, Ruolin Wang, Dan Liu, Tong Zhang, Sanyuan Ma, Ling Jia
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引用次数: 0

摘要

昆虫变态是一个复杂的内分泌级联反应,由幼体激素(JH)和20-羟基蜕皮激素(20E)之间的动态相互作用所控制。虽然JH已被确定为一种关键的发育调节因子,但其细胞内信号传导机制仍不完整。在本研究中,我们在鳞翅目模型家蚕中实施了全基因组CRISPR筛选策略,以系统地识别JH信号的组成部分。首先,我们开发了一种jh敏感启动子控制下EGFP表达的jh响应报告细胞系,能够实时监测途径激活情况。然后,我们建立了针对家蚕细胞的基因组级CRISPR敲除文库,并结合高通量流式细胞术和下一代测序技术进行筛选。系统分析显示,共有387个候选基因显著调节JH信号活性(231个富集,156个缺失)。功能注释聚类显示主要富集于膜生物发生过程和小分子代谢。值得注意的是,通过整合基因组筛选和实验验证,我们发现酪氨酸激酶受体BmALK是JH信号转导的重要调控节点。本研究首次建立了全代谢昆虫JH信号的综合遗传图谱,为JH信号通路的功能挖掘和鳞翅目JH信号转导的膜相关分子证据提供了强有力的平台。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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.

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来源期刊
Insect Science
Insect Science 生物-昆虫学
CiteScore
7.80
自引率
5.00%
发文量
1379
审稿时长
6.0 months
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.
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