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Feeding microRNA mimics and inhibitors to study the mechanisms of insect miRNA regulation and development
Helicoverpa armigera , a kind of insect of the genus Noctuidae, is an important drill-bit pest of the cotton bud, mainly feeding on buds, flowers, bells, and feeding young leaves. MicroRNAs are a class of non-coding small RNAs (18-25 nt) that play important roles in various biological processes, including development and gene regulation. Researchers at the University of Queensland in Australia have systematically studied the regulation of the development and reproduction of miRNAs specific to Helicoverpa armigera by synthetic miRNA mimics and inhibitors*. The results were published in the recent Insect Biochem Mol Biol .
First, the researchers identified five miRNAs specifically expressed in the larval stage by high-throughput miRNA sequencing of the larvae of the cotton bollworm (Lianchuan Biotechnology), and selected har-miR-2002b as a Northern hybrid analysis. The target of the study, using the biological software to predict its target gene is Trypsin-like Serine Protease (Ha-TLP). Subsequently, using the synthetic inhibitor and mimic to feed the cotton bollworm, it was found that inhibitor and mimic could successfully inhibit or overexpress har-MIR-2002 in the larvae of Helicoverpa armigera. Biological assays showed that under the condition of feeding har-miR-2002b mimic, 70% of the larvae had reduced fertility and 40% of the larvae died; most of the larvae became malformed and the moth rate was extremely low. It was confirmed by RT-PCR that Ha-TLP is a target gene of har-miR-2002b. In vivo and in vitro studies showed that Ha-TLP transcript/protein levels and enzyme activity were significantly reduced by miRNA mimic; in contrast, Ha-TLP transcripts were significantly up-regulated by har-miR-2002b inhibitor.
The above studies indicate that miRNA mimic or inhibitor can be used to inhibit or overexpress miRNA in vivo and can be used for the study of miRNA follow-up functions.
*Jayachandran B, Hussain M, Asgari S. 2012. An insect trypsin-like serine protease as a target of microRNA: Utilization of microRNA mimics and inhibitors by oral feeding. Insect Biochem Mol Biol [Epub ahead of print].
Further reading: JVI: Insects use siRNA instead of miRNA to protect against virus infection
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