Cytochrome P450 monooxygenases (CYPs) play essential physiological functions in insects. CYP303A1 is highly conserved in insect species studied to date, and shows an indispensable role for adult eclosion in both Locusta migratoria and Drosophila melanogaster. However, how CYP303A1 is regulated to control insect developmental processes remains uninvestigated. In this study, we discovered functional binding sites for miR-184 in the coding sequence of LmCYP303A1. The luciferase reporter assay showed that miR-184 could target LmCYP303A1 and regulate its expression in vitro. Furthermore, overexpression of miR-184 through microinjection of agomir to locusts reduced the transcripts of LmCYP303A1 and led to abnormal molting, which is similar to the phenotype of silencing LmCYP303A1 by direct injection of dsLmCYP303A1 to locusts. Meanwhile, down-regulation of miR-184 by injection of antagomir increased the LmCYP303A1 transcript and caused molting defects. These findings suggested that miR-184 could target LmCYP303A1 to regulate the molting process in L. migratoria, which might be considered as a novel target for pest control.

中文翻译：

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microRNA miR-184 调节 CYP303A1 转录水平以控制蝗虫的蜕皮

细胞色素 P450 单加氧酶 (CYPs) 在昆虫中发挥重要的生理功能。CYP303A1在迄今为止研究的昆虫物种中高度保守，并且在Locusta migratoria和Drosophila melanogaster 中对成虫羽化显示出不可或缺的作用。然而，如何调节CYP303A1以控制昆虫发育过程仍未得到研究。在这项研究中，我们在LmCYP303A1的编码序列中发现了 miR-184 的功能结合位点。荧光素酶报告基因检测表明 miR-184 可以靶向LmCYP303A1并在体外调节其表达. 此外，的miR-184的过表达通过agomir蝗虫的显微注射减少的转录物LmCYP303A1并导致异常蜕皮，其类似于沉默的表型LmCYP303A1通过直接注射DS的LmCYP303A1蝗虫。同时，通过注射antagomir下调miR-184会增加LmCYP303A1转录本并导致蜕皮缺陷。这些发现表明 miR-184 可以靶向LmCYP303A1来调节L. migratoria的蜕皮过程，这可能被认为是害虫防治的新靶点。