Abstract
BackgroundmicroRNAs (miRNAs) are small non-coding RNAs widely identified in many mosquitoes. They are reported to play important roles in development, differentiation and innate immunity. However, miRNAs in Anopheles sinensis, one of the Chinese malaria mosquitoes, remain largely unknown.MethodsWe investigated the global miRNA expression profile of An. sinensis using Illumina Hiseq 2000 sequencing. Meanwhile, we applied a bioinformatic approach to identify potential miRNAs in An. sinensis. The identified miRNA profiles were compared and analyzed by two approaches. The selected miRNAs from the sequencing result and the bioinformatic approach were confirmed with qRT-PCR. Moreover, target prediction, GO annotation and pathway analysis were carried out to understand the role of miRNAs in An. sinensis.ResultsWe identified 49 conserved miRNAs and 12 novel miRNAs by next-generation high-throughput sequencing technology. In contrast, 43 miRNAs were predicted by the bioinformatic approach, of which two were assigned as novel. Comparative analysis of miRNA profiles by two approaches showed that 21 miRNAs were shared between them. Twelve novel miRNAs did not match any known miRNAs of any organism, indicating that they are possibly species-specific. Forty miRNAs were found in many mosquito species, indicating that these miRNAs are evolutionally conserved and may have critical roles in the process of life. Both the selected known and novel miRNAs (asi-miR-281, asi-miR-184, asi-miR-14, asi-miR-nov5, asi-miR-nov4, asi-miR-9383, and asi-miR-2a) could be detected by quantitative real-time PCR (qRT-PCR) in the sequenced sample, and the expression patterns of these miRNAs measured by qRT-PCR were in concordance with the original miRNA sequencing data. The predicted targets for the known and the novel miRNAs covered many important biological roles and pathways indicating the diversity of miRNA functions. We also found 21 conserved miRNAs and eight counterparts of target immune pathway genes in An. sinensis based on the analysis of An. gambiae.ConclusionsOur results provide the first lead to the elucidation of the miRNA profile in An. sinensis. Unveiling the roles of mosquito miRNAs will undoubtedly lead to a better understanding of mosquito biology and mosquito-pathogen interactions. This work lays the foundation for the further functional study of An. sinensis miRNAs and will facilitate their application in vector control.
Highlights
MicroRNAs are small non-coding RNAs widely identified in many mosquitoes
In conclusion, our study provides a comprehensive account of the miRNA profile of An. sinensis by combining the deep sequencing and the bioinformatic approach
Efforts were undertaken to understand the targets of miRNAs which can provide a better understanding of their biological function in mosquito biology and immunity and provide implications for effective control in the future
Summary
MicroRNAs (miRNAs) are small non-coding RNAs widely identified in many mosquitoes. They are reported to play important roles in development, differentiation and innate immunity. MiRNAs in Anopheles sinensis, one of the Chinese malaria mosquitoes, remain largely unknown. Indigenous malaria in some places, together with the great challenge posed by imported Plasmodium falciparum malaria [3], would definitely hinder the process of elimination in the coming years. One of the important malaria vectors in China, is still considered as the primary vector of P. vivax malaria due to its wide distribution and high density. Malaria used to be prevalent in most parts of the flatlands in China between 25 and 33° N where this species was incriminated as the most dominant malaria vector [5]. Effective integrated vector control research is needed to sustain the gains achieved so far and to achieve global malaria elimination in the future
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