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Insect Biochem Mol Biol

Title:		piggyBac-based transgenic RNAi of serine protease 2 results in male sterility in Hyphantria cunea
Author(s):		Li X; Liu Q; Bi H; Wang Y; Xu X; Sun W; Zhang Z; Huang Y;
Address:		"Laboratory of Evolutionary and Functional Genomics, School of Life Sciences, Chongqing University, Chongqing, 401331, China; CAS Key Laboratory of Insect Developmental and Evolutionary Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai, 200030, China. CAS Key Laboratory of Insect Developmental and Evolutionary Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai, 200030, China. Institute of Sericulture and Tea Research, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China. Laboratory of Evolutionary and Functional Genomics, School of Life Sciences, Chongqing University, Chongqing, 401331, China. Laboratory of Evolutionary and Functional Genomics, School of Life Sciences, Chongqing University, Chongqing, 401331, China. Electronic address: zezhang@cqu.edu.cn. CAS Key Laboratory of Insect Developmental and Evolutionary Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai, 200030, China; CAS Center for Excellence in Biotic Interactions, University of Chinese Academy of Sciences, Beijing, 100049, China. Electronic address: yphuang@sibs.ac.cn"
Journal Title:		Insect Biochem Mol Biol
Year:		2022
Volume:		20220205
Issue:
Page Number:		103726 -
DOI:		10.1016/j.ibmb.2022.103726
ISSN/ISBN:		1879-0240 (Electronic) 0965-1748 (Linking)
Abstract:		"Fall webworm, Hyphantria cunea, is a global invasive forest pest that causes serious damage to the economy and ecosystem of agriculture and forestry. Due to the extent of the problem and the difficulty of conventional chemical control, new technologies must be pursued, such as genetic-based inheritable insect sterile technology (gSIT), which exhibits promise for pest control. In the present study, we established a piggyBac-based transgenic system in fall webworm and generated a dominant male-sterile strain by targeting the seminal fluid protein serine protease 2 (Hcser2), displaying an outstanding trait of gSIT. First, an RNA polymerase type III (Pol III) promoter, the HcU62 small nuclear RNA (snRNA) gene promoter, was identified and characterized through direct injection of RNAi plasmids in vivo. Quantitative real-time PCR revealed that HcU62 had the greatest knockdown efficiency of the Hcyellow gene among five short hairpin RNA (shRNA) plasmids tested, designated HcU61-HcU65. Second, subsequent application of piggyBac-based transgenic RNAi (HcU62: shHcyellow, Ysh2) significantly reduced the expression level of the Hcyellow gene, resulting in a stable yellow observable phenotype from the larval to pupal stages in Ysh2 transgenic mutants. Finally, an HcU62-driven transgenic RNAi strain targeting the Hcser2 gene was obtained, resulting in a dominant male-sterile phenotype. Significantly, this process did not affect the growth, development, mating behavior or egg laying of the mutants, and the dominant sterile trait could be inherited in the next generation through female Hcser2 mutants. Furthermore, CRISPR/Cas9-mediated disruption of the Hcser2 gene further confirmed the dominant sterile phenotype, supporting it as a generalized target for genetic control of H. cunea. This study reports the first piggyBac-mediated transgenic system in H. cunea, providing a promising genetic method for controlling this pest by targeting Hcser2 gene"
Keywords:		"Animals Ecosystem Female *Infertility, Male/genetics Male *Moths/genetics RNA Interference Serine Proteases/genetics CRISPR/Cas9 Hyphantria cunea Male sterility piggyBac ser2;"
Notes:		"MedlineLi, Xiaowei Liu, Qun Bi, Honglun Wang, Yaohui Xu, Xia Sun, Wei Zhang, Ze Huang, Yongping eng Research Support, Non-U.S. Gov't England 2022/02/09 Insect Biochem Mol Biol. 2022 Apr; 143:103726. doi: 10.1016/j.ibmb.2022.103726. Epub 2022 Feb 5"