CRISPR/Cas9-mediated editing of the melanization gene ebony in the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata
文献类型： 外文期刊
作者： Chen, Shimin 1 ; Ruan, Yalin 1 ; Li, Zhaoyang 1 ; Zhou, Chuqiao 1 ; Chen, Qi 1 ; Zhou, Xuguo 2 ; Zhang, Youjun 3 ; Yang, Chunxiao 4 ; Pan, Huipeng 1 ;
作者机构： 1.South China Agr Univ, Integrat Microbiol Res Ctr, State Key Lab Conservat & Utilizat Subtrop Agrobio, Minist Educ, Guangzhou 510642, Peoples R China
2.Univ Illinois Champaign Urbana, Dept Entomol, Urbana, IL 61801 USA
3.Chinese Acad Agr Sci, Dept Plant Protect, Inst Vegetables & Flowers, Beijing 100081, Peoples R China
4.South China Agr Univ, State Key Lab Conservat & Utilizat Subtrop Agrobio, Guangzhou 510642, Peoples R China
关键词： CRISPR/Cas9; RNAi; Ebony; Cuticle melanization; Henosepilachna vigintioctopunctata
期刊名称：PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY （ 影响因子：4.0； 五年影响因子：4.5 ）
ISSN： 0048-3575
年卷期： 2025 年 208 卷
页码：
收录情况： SCI
摘要： The melanization process, which is essential for the proper functioning of the cuticle, has been extensively investigated for its enzymatic roles and physiological effects. Henosepilachna vigintioctopunctata, a significant pest species, presents considerable economic threats. However, due to the variable efficiency of RNA interference for genetic manipulation, establishing a CRISPR/Cas9 system is crucial for providing a more precise and reliable method for functional genomics in this non-model insect. In this study, we first utilized RNAi to investigate Hvebony, which encodes N-(3-alanyldopamine, a critical compound in cuticle melanization. Subsequently, we introduced CRISPR/Cas9 for the first time in H. vigintioctopunctata. RNAi experiments revealed that knockdown of Hvebony resulted in abnormal melanin accumulation and low mortality rates, indicating its involvement in cuticle tanning. A novel CRISPR/Cas9 workflow was established, successfully resulting in the knocking out of Hvebony and the creation of a stable mutant strain characterized by dark pigmentation and low fitness costs. This study establishes Hvebony as a promising molecular marker for genetic studies in H. vigintioctopunctata. Moreover, it can be utilized in the development of genome editing control strategies and for analyses of gene function in H. vigintioctopunctata.
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