Functional analysis of the BmRpt4 gene during wing development in the silkworm, Bombyx mori
Author of the article:SUN Xia；ZHANG Jing ；YIN Jin；QIN Sheng； ZHANG GuoZheng；LI MuWang
Author's Workplace：1. College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212018, China; 2. The Sericultural Research Institute, Chinse Academy of Agricultural Sciences, Zhenjiang 212018, China
Key Words：minute wing, proteasome, CRISPR/Cas9, genomic editing, silkworm
Abstract：[Objectives] To study and analyze the function of the BmRpt4 gene (Regulatory particle triple-A ATPase 4, BGIBMGA010794) during wing development in the silkworm Bombyx mori. [Methods] BmRpt4 gRNAs and Cas9 mRNA were directly injected into silkworm embryos using the CRISPR/Cas9 genome editing technique. Phenotypes of G0 adults were observed and the proportion of adults with minute wings was statistically analyzed. Mutation of BmRpt4 was detected in adults with minute wings. [Results] All G0 adults in the control group had normal wings whereas 66.67% of G0 adults in the experimental group had minute wings. The BmRpt4 gene was deleted, or inserted, in different length fragments at the two gRNAs locii in G0 individuals with minute wings. These results indicate that individuals developed minute wings due to knock-down of the BmRpt4 gene. [Conclusion] The BmRpt4 gene plays an important role in wing development in the silkworm, and in regulating proteasome in the development of biological tissues and organs.