MicroRNA-14 regulates larval development time in Bombyx mori. (February 2018)

• Record Type: Journal Article
• Authors: Liu, Zulian; Ling, Lin; Xu, Jun; Zeng, Baosheng; Huang, Yongping; Shang, Peng; Tan, Anjiang
• Is Part Of: Insect biochemistry and molecular biology. Volume 93(2018)
• Page Start: 57
• Page End: 65
• Publication Date: 2018-02
• Subjects: Bombyx mori -- miR-14 -- Ecdysone -- Gal4/UAS -- CRISPR/Cas9; Insect biochemistry -- Periodicals; Insects -- Physiology -- Periodicals; Insects -- Molecular aspects -- Periodicals; Biochemistry -- Periodicals; Insectes -- Biochimie -- Périodiques; Insectes -- Composition -- Périodiques; Insectes -- Physiologie -- Périodiques; Insectes -- Aspect moléculaire -- Périodiques; Biochimie -- Périodiques; Biochemistry; Insect biochemistry; Insects -- Molecular aspects; Insects -- Physiology; Periodicals; 572.8157
• DOI: 10.1016/j.ibmb.2017.12.009
• Languages: English
• ISSNs: 0965-1748

Abstract

Abstract: MicroRNAs (miRNA) regulate multiple physiological processes including development and metamorphosis in insects. In the current study, we demonstrate that a conserved invertebrate miRNA-14 (miR-14) plays an important role in ecdysteroid regulated development in the silkworm Bombyx mori, a lepidopteran model insect. Ubiquitous transgenic overexpression of miR-14 using the GAL4/UAS system resulted in delayed silkworm larval development and smaller body size of larva and pupa with decrease in ecdysteriod titers. On the contrary, miR-14 disruption using the transgenic CRISPR/Cas9 system led to a precocious wandering stage with increase in ecdysteriod titers. We identified that the hormone receptor E75 ( E75 ) and the ecdysone receptor isoform B ( ECR-B ), which both serve as essential mediators in the ecdysone signaling pathway, as putative target genes of miR-14 by in silico target prediction. Dual-luciferase reporter assays confirmed the binding of miR-14 to the 3′UTRs of E75 and ECR-B in a mammalian HEK293T cell line. Furthermore, transcription levels of E75 and ECR-B were significantly affected in both miR-14 overexpression and knockout transgenic animals. Taken together, our data suggested that the canonical invertebrate miR-14 is a general regulator in maintaining ecdysone homeostasis for normal development and metamorphosis in B. mori . Graphical abstract: Highlights: The conserved invertebrate miR-14 plays a role in ecdysteroid regulated development in Bombyx mori. Ubiquitous transgenic overexpression of miR-14 using the GAL4/UAS system delayed larval development. miR-14 disruption using the transgenic CRISPR/Cas9 system led to a precocious wandering stage. E75 and EcR-B are putative target genes of miR-14 in B. mori.