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Study on the function of Helicoverpa armigera Wnt1 gene using CRISPR/Cas9 system
Institution:1. Department of Plant Protection, Henan Agricultural University, Zhengzhou 450000, China;2. Key Laboratory of New Pesticide Development and Application, Henan Agricultural University, Zhengzhou 450000, China;3. Green Pesticide Creation Engineering Technology Research Center, Henan Agricultural University, Zhengzhou 450000, China
Abstract:The Wnt signaling pathway, as a highly conserved signaling pathway in evolution, plays an important role in many biological processes. The research of Wnt signaling pathway through gene editing has been implemented in a variety of organisms. Among the various genome editing tools available for functional genomic research, CRISPR is popular because of its ease of use and versatility. Here, we use the CRISPR/Cas9 system to knock out the HaWnt1 gene of the important agricultural pest Helicoverpa armigera to explore the impacts on embryo development. Direct injection of Cas9 protein and Wnt1-specific single guide RNA (sgRNA) into H. armigera embryos successfully induced Wnt1 gene deletion mutants, which showed high lethality, abnormal segmentation, defected appendages and defected pigmentation. qRT-PCR analysis revealed that the deletion of Wnt1 gene affected the expression of several genes, which were closely related to the growth and development of insects. Our results indicate that HaWnt1 signaling pathway is essential for embryonic development of H. armigera. The study of the function of HaWnt1 gene not only lays the foundation for the study of the somatic development pattern of H. armigera, but also provides a candidate gene for genetic control of H. armigera.
Keywords:Wnt1  CRISPR/Cas9 system  Genome editing  Embryos
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