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The temporal patterns of miRNA-target interactions obtained in this study provide new insights into the roles of miRNAs in tightly controlled gene expression associated with blood-feeding and mosquito oogenesis.Ĭitation: Fu X, Liu P, Dimopoulos G, Zhu J (2020) Dynamic miRNA-mRNA interactions coordinate gene expression in adult Anopheles gambiae. Drastic changes in mRNA abundance have been previously reported when adult female mosquitoes attend to varying nutritional sources and physiological demands. Interestingly, many mosquito miRNAs exhibit flexibility to use distinct sequences at different stages to pair with their targets, greatly altering target selectivity and expanding target repertoire of miRNAs. Analyses of the target sequences reveal how individual miRNAs accomplish their target recognition in mosquitoes. Using an experimental approach, this study captures thousands of endogenous miRNA-target interactions in female mosquitoes at several critical stages during adult reproduction. Target identification is even more difficult in mosquitoes where miRNA-mRNA pairing rules are poorly characterized. Thus, identification of authentic miRNA targets remains a big challenge. The interactions between miRNAs and target RNAs are generally stage-specific and context-dependent. Metazoan miRNAs typically bind to partially complementary sites in their target mRNAs. It also provides a roadmap for additional studies focused on regulatory functions of miRNAs in Anopheles gambiae. The global mapping of miRNA-target interactions contributes to our understanding of miRNA targeting specificity in non-model organisms. Furthermore, many miRNA-mRNA interactions displayed stage-specific patterns, especially for those genes involved in metabolism, suggesting that miRNAs play critical roles in precise control of gene expression to cope with enormous physiological demands associated with egg production. Interestingly, some miRNAs exhibited flexibility to use distinct sequences at different stages for target recognition. The time-lapse study of miRNA-target interactomes in adult female mosquitoes revealed dynamic miRNA regulation of gene expression in response to varying nutritional sources and physiological demands. Enriched motifs in the diverse targets of each miRNA indicated that the majority of mosquito miRNAs rely on seed-based canonical target recognition, while noncanonical miRNA binding sites are widespread and often contain motifs complementary to the central or 3’ ends of miRNAs. The miRNA-mRNA interaction dataset provided an invaluable opportunity to decipher targeting rules of mosquito miRNAs.
Using two interactions detected in this manner, miR-309- SIX4 and let-7- kr-h1, we demonstrated the reliability of this experimental approach in identifying in vivo gene regulation by miRNAs. Thousands of miRNA-target pairs were captured after direct ligation of the miRNA and its cognate target transcript in endogenous Argonaute–miRNA–mRNA complexes. In this study, we used CLEAR-CLIP assays to systematically analyze miRNA-mRNA interactions in adult female Anopheles gambiae mosquitoes. Identification of bona fide targets remains the bottleneck for functional studies of miRNAs.
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MicroRNAs (miRNAs) are increasingly recognized as important regulators of many biological processes in mosquitoes, vectors of numerous devastating infectious diseases.