MicroRNA Let-7 regulates molting and metamorphosis in the silkworm, Bombyx mori
MicroRNA Let-7 regulates molting and metamorphosis in the silkworm, Bombyx mori
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MicroRNA Let-7 调节家蚕 (Bombyx mori) 的蜕皮和变态。
DOI：
10.1016/j.ibmb.2014.06.011
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发表时间：
2014-10-01
影响因子：
3.8
通讯作者：
Tan, Anjiang
中科院分区：
文献类型：
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作者：
Ling, Lin;Ge, Xie;Tan, Anjiang
MicroRNAs(MiRNAs)是一类内源性的、非编码的、调节性的RNA分子，它通过与mRNA靶标的3‘UTRs结合，在转录后调节基因的表达，从而导致它们的降解或翻译抑制。在昆虫中，miRNAs在各种生物学过程中的重要作用已经在果蝇身上被证明。然而，由于有限的遗传工具，miRNAs在大多数昆虫物种中的生物学作用几乎没有被揭示。在本研究中，我们将转基因miRNA海绵(miR-SP)技术与家蚕GAL4/UAS双元系统相结合，以开发进化保守的miRNA let-7的生物学功能。我们成功地建立了转基因家蚕品系，其中针对Bmlet-7种子区的miRNA海绵构建物在3-GAL4驱动程序的指导下以普遍的方式表达。转基因动物Bmlet-7的表达降低，导致幼虫-幼虫和幼虫-幼虫过渡期间的发育停滞。同时，预测的Bmlet-7靶基因FTZ-F1和Eip74EF(E74)的表达水平在转基因动物中升高，这两个基因是蜕皮激素途径的关键调控因子。本研究是转基因miR-SP技术在非果蝇昆虫中应用的首次报道，这不仅有助于更好地了解let-7的生物学作用，而且为未来昆虫miRNA的功能分析提供了极大的便利。(C)2014爱思唯尔有限公司。保留所有权利。
MicroRNAs (miRNAs) are a class of endogenous, non-coding, regulatory RNA molecules that post-transcriptionally regulate gene expression by binding to the 3'UTRs of mRNA targets and thus cause their degradation or translational inhibition. In insects, important roles of miRNAs in various biological processes have been demonstrated in Drosophila melanogaster. However, biological roles of miRNAs are barely unveiled in the majority of insect species due to limited genetic tools. In the present study, we introduce the transgenic miRNA sponge (miR-SP) technology combining with the binary GAL4/UAS system in the domesticated silkworm, Bombyx mori, to exploit the biological function of an evolutionally conserved miRNA, let-7. We successfully established transgenic silkworm lines in which a miRNA sponge construct targeting BmLet-7 seed region was expressed in a ubiquitous manner directed by A3-GAL4 driver. Transgenic animals showed decreased expression of BmLet-7, leading to developmental arrestment during the larval-larval and larval-pupal transition. Simultaneously, expression levels of the predicted BmLet-7 target genes, FTZ-F1 and Eip74EF (E74), key regulatory factors in the ecdysone pathway, were elevated in transgenic animals. The current study is the first report on application of the transgenic miR-SP technology in non-drosophilid insects, which will not only contribute to better understanding of let-7 biological roles, but also greatly facilitate future miRNA functional analysis in insects. (C) 2014 Elsevier Ltd. All rights reserved.