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

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

机构地区：[1]State Key Laboratory of Silkworm Genome Biology,Southwest University,Chongqing,China [2]Biological Science Research Center,Southwest University,Chongqing,China

出　　处：《Insect Science》2024年第1期47-58,共12页昆虫科学（英文版）

基　　金：This work was supported by grants from the National Natural Science Foundation of China(No.32122084);Chongqing Natural Science Foundation(No.cstc2021ycjh-bgzxm0005);PhD Start-Up Foundation of Southwest University(No.SWU120012);Fundamental Research Funds for the Central Universities(No.SWU-KT22042).None of these fundings played any role in the design of the study,collection,analysis,or interpretation of data or in the writing of the manuscript.

摘　　要：Metamorphosis is a complex developmental process involving multiple pathways 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 understood.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 successfully 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 nutrition 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.

关 键 词：CRISPR/Cas9 CRISPR library screening ECDYSONE 20E signal transduction SILKWORM whole genome editing 

分 类 号：Q78[生物学—分子生物学]