MiR-2 family targets awd and fng to regulate wing morphogenesis in Bombyx mori
MiR-2 family targets awd and fng to regulate wing morphogenesis in Bombyx mori
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DOI：
10.1080/15476286.2015.1048957
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发表时间：
2015-07-03
期刊：
影响因子：
4.1
通讯作者：
Tan, Anjiang
中科院分区：
文献类型：
--
作者：
Ling, Lin;Ge, Xie;Tan, Anjiang
microRNAs（miRNAs）是一种转录后调节因子，能够靶向特定的mRNA进行抑制，在昆虫翅的形态发生等生物学过程中发挥重要作用。miR-2是一个无脊椎动物特异性的miRNA家族，在果蝇中被预测参与调节Notch信号通路。我们发现miR-2在鳞翅目模式昆虫家蚕的翅形态发生中起着至关重要的作用。使用Gal 4/UAS系统转基因过表达miR-2簇导致变形的成虫翅膀，支持miR-2调节正常翅膀形态发生所必需的功能的结论。两个基因，异常翅盘（awd）和边缘（fng），这是在Notch信号的正调控，被确定为miR-2的目标，并通过双荧光素酶报告分析验证。在转基因动物中，awd和fng表达产物的相对丰度都显著降低，表明它们与翅表型异常有关。此外，使用CRISPR/Cas9系统和敲除突变体对awd和fng进行的体细胞诱变分析也导致了与在miR-2过表达转基因动物中观察到的类似的变形翅膀。miR-2在小菜蛾翅形态发生中的关键作用可能为未来鳞翅目害虫防治提供潜在靶点。
MicroRNAs (miRNAs) are post-transcriptional regulators that target specific mRNAs for repression and thus play key roles in many biological processes, including insect wing morphogenesis. miR-2 is an invertebrate-specific miRNA family that has been predicted in the fruit fly, Drosophila melanogaster, to be involved in regulating the Notch signaling pathway. We show here that miR-2 plays a critical role in wing morphogenesis in the silkworm, Bombyx mori, a lepidopteran model insect. Transgenic over-expression of a miR-2 cluster using a Gal4/UAS system results in deformed adult wings, supporting the conclusion that miR-2 regulates functions essential for normal wing morphogenesis. Two genes, abnormal wing disc (awd) and fringe (fng), which are positive regulators in Notch signaling, are identified as miR-2 targets and validated by a dual-luciferase reporter assay. The relative abundance of both awd and fng expression products was reduced significantly in transgenic animals, implicating them in the abnormal wing phenotype. Furthermore, somatic mutagenesis analysis of awd and fng using the CRISPR/Cas9 system and knock-out mutants also resulted in deformed wings similar to those observed in the miR-2 overexpression transgenic animals. The critical role of miR-2 in Bombyx wing morphogenesis may provide a potential target in future lepidopteran pest control.