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MicroRNA-14 regulates larval development time in Bombyx mori
Zulian Liu1, Lin Ling2, Jun Xu2
1Faculty of Life Sciences, Northwestern Polytechnical University, Xi'an 710072, China; Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Insect Biochemistry and Molecular Biology
|December 31, 2017
Summary
MicroRNA-14 (miR-14) is crucial for silkworm development. It regulates ecdysteroid levels, impacting larval growth and metamorphosis by targeting key hormone receptors.
Area of Science:
- Developmental Biology
- Molecular Biology
- Entomology
Background:
- MicroRNAs (miRNAs) are key regulators of gene expression in eukaryotes.
- Insect development and metamorphosis are tightly controlled by hormones like ecdysteroids.
- The silkworm Bombyx mori is a valuable model organism for studying insect development.
Purpose of the Study:
- To investigate the role of the conserved invertebrate microRNA-14 (miR-14) in ecdysteroid-regulated development in Bombyx mori.
- To identify the molecular targets of miR-14 within the ecdysone signaling pathway.
Main Methods:
- Transgenic overexpression of miR-14 using the GAL4/UAS system.
- CRISPR/Cas9-mediated knockout of miR-14.
- Measurement of ecdysteroid titers.
- In silico prediction of miRNA targets.
- Dual-luciferase reporter assays in HEK293T cells.
- Analysis of target gene transcription levels.
Main Results:
- Overexpression of miR-14 led to delayed larval development, smaller body size, and decreased ecdysteroid titers.
- miR-14 disruption resulted in precocious wandering and increased ecdysteroid titers.
- Ecdysone receptor 75 (E75) and ecdysone receptor isoform B (ECR-B) were identified as direct targets of miR-14.
- miR-14 directly binds to the 3'UTRs of E75 and ECR-B, affecting their transcription.
Conclusions:
- The invertebrate miR-14 acts as a critical regulator of ecdysone homeostasis in Bombyx mori.
- miR-14 is essential for normal development and metamorphosis by modulating the ecdysone signaling pathway.
- These findings highlight miR-14 as a conserved regulator in insect endocrine signaling.
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