首页|Genome-wide CRISPR screening reveals key genes and pathways associated with 20-hydroxyecdysone signal transduction in the silkworm(Bombyx mori)

Genome-wide CRISPR screening reveals key genes and pathways associated with 20-hydroxyecdysone signal transduction in the silkworm(Bombyx mori)

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Metamorphosis is a complex developmental process involving multiple path-ways and a large number of genes that are regulated by juvenile hormone(JH)and 20-hydroxyecdysone(20E).Despite important progress in understanding various aspects of silkworm biology,the hormone signaling pathway in the silkworm remains poorly under-stood.Genome-wide screening using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-based libraries has recently emerged as a novel method for analyzing genome function,enabling further research into essential genes,drug targets,and virus-host interaction.Previously,we constructed a genome-wide CRISPR/Cas9-based library of the silkworm(Bombyx mori)and success-fully revealed the genes involved in biotic or abiotic stress factor responses.In this study,we used our silkworm CRISPR library and large-scale genome-wide screening to analyze the key genes in the silkworm 20E signaling pathway and their mechanisms of action.Functional annotation showed that 20E regulates key proteins in processes that mainly occur in the cytoplasm and nucleus.Pathway enrichment analysis showed that 20E can activate phosphorylation and may affect innate immunity,interfere with intracellular nu-trition and energy metabolism,and eventually cause cell apoptosis.The screening results were experimentally validated by generating cells with knockout alleles of the relevant genes,which had increased tolerance to 20E.Our findings provide a panoramic overview of signaling in response to 20E in the silkworm,underscoring the utility of genome-wide CRISPR mutant libraries in deciphering hormone signaling pathways and the mechanisms that regulate metamorphosis in insects.

CRISPR/Cas9CRISPR library screeningecdysone20E signal transduc-tionsilkwormwhole genome editing

Hao Sun、Jingya Chen、Ruolin Wang、Dan Liu、Na Zhang、Tong Zhang、Ling Jia、Sanyuan Ma、Qingyou Xia

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State Key Laboratory of Silkworm Genome Biology,Southwest University,Chongqing,China

Biological Science Research Center,Southwest University,Chongqing,China

National Natural Science Foundation of ChinaChongqing Natural Science FoundationPhD Start-Up Foundation of Southwest UniversityFundamental Research Funds for the Central Universities

32122084cstc2021 ycjhbgzxm0005SWU120012SWU-KT22042

2024

10.1111/1744-7917.13237

中国昆虫科学(英文版)
中国昆虫学会 中科院动物所

中国昆虫科学(英文版)

CSTPCD
影响因子：0.484
ISSN：1672-9609
年,卷(期)：2024.31(1)
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