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首页> 外文期刊>BMC Genomics >Direct sequencing and expression analysis of a large number of miRNAs in Aedes aegypti and a multi-species survey of novel mosquito miRNAs
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Direct sequencing and expression analysis of a large number of miRNAs in Aedes aegypti and a multi-species survey of novel mosquito miRNAs

机译：埃及伊蚊中大量miRNA的直接测序和表达分析以及新型蚊子miRNA的多物种调查

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Background MicroRNAs (miRNAs) are a novel class of gene regulators whose biogenesis involves hairpin structures called precursor miRNAs, or pre-miRNAs. A pre-miRNA is processed to make a miRNA:miRNA* duplex, which is then separated to generate a mature miRNA and a miRNA*. The mature miRNAs play key regulatory roles during embryonic development as well as other cellular processes. They are also implicated in control of viral infection as well as innate immunity. Direct experimental evidence for mosquito miRNAs has been recently reported in anopheline mosquitoes based on small-scale cloning efforts. Results We obtained approximately 130, 000 small RNA sequences from the yellow fever mosquito, Aedes aegypti, by 454 sequencing of samples that were isolated from mixed-age embryos and midguts from sugar-fed and blood-fed females, respectively. We also performed bioinformatics analysis on the Ae. aegypti genome assembly to identify evidence for additional miRNAs. The combination of these approaches uncovered 98 different pre-miRNAs in Ae. aegypti which could produce 86 distinct miRNAs. Thirteen miRNAs, including eight novel miRNAs identified in this study, are currently only found in mosquitoes. We also identified five potential revisions to previously annotated miRNAs at the miRNA termini, two cases of highly abundant miRNA* sequences, 14 miRNA clusters, and 17 cases where more than one pre-miRNA hairpin produces the same or highly similar mature miRNAs. A number of miRNAs showed higher levels in midgut from blood-fed female than that from sugar-fed female, which was confirmed by northern blots on two of these miRNAs. Northern blots also revealed several miRNAs that showed stage-specific expression. Detailed expression analysis of eight of the 13 mosquito-specific miRNAs in four divergent mosquito genera identified cases of clearly conserved expression patterns and obvious differences. Four of the 13 miRNAs are specific to certain lineage(s) within mosquitoes. Conclusion This study provides the first systematic analysis of miRNAs in Ae. aegypti and offers a substantially expanded list of miRNAs for all mosquitoes. New insights were gained on the evolution of conserved and lineage-specific miRNAs in mosquitoes. The expression profiles of a few miRNAs suggest stage-specific functions and functions related to embryonic development or blood feeding. A better understanding of the functions of these miRNAs will offer new insights in mosquito biology and may lead to novel approaches to combat mosquito-borne infectious diseases.
机译：背景技术MicroRNA（miRNA）是一类新型的基因调节剂，其生物发生涉及称为前体miRNA或pre-miRNA的发夹结构。将pre-miRNA加工成miRNA：miRNA *双链体，然后将其分离以生成成熟的miRNA和miRNA *。成熟的miRNA在胚胎发育以及其他细胞过程中起着关键的调节作用。它们还涉及病毒感染的控制以及先天免疫。基于小规模克隆的努力，最近在按蚊的蚊子中报道了蚊子miRNA的直接实验证据。结果通过对分别来自混合年龄胚胎和食糖雌性和混血雌性的混合年龄胚胎和中肠分离的样品进行454测序，我们从黄热蚊，埃及伊蚊获得了大约130、000个小RNA序列。我们还对Ae进行了生物信息学分析。 aegypti基因组装配，以鉴定其他miRNA的证据。这些方法的结合在Ae中发现了98种不同的pre-miRNA。 aegypti可以产生86种不同的miRNA。目前仅在蚊子中发现了十三种miRNA，包括在这项研究中鉴定的八种新型miRNA。我们还确定了对miRNA末端先前注释的miRNA的五个潜在修订，两个高度丰富的miRNA *序列案例，14个miRNA簇以及17个案例，其中不止一个pre-miRNA发夹产生相同或高度相似的成熟miRNA。血源雌性中肠中的许多miRNA含量高于糖源雌性中肠中的水平，这在其中两个miRNA的Northern印迹分析中得到了证实。 Northern印迹法还揭示了几种miRNA，它们显示了阶段特异性表达。在四个不同的蚊子属中的13个蚊子特异性miRNA中的八个进行了详细的表达分析，确定了明确保守的表达模式和明显差异的病例。 13种miRNA中有4种是特定于蚊子中某些谱系的。结论本研究首次对Ae中的miRNA进行了系统分析。 aegypti，并为所有蚊子提供了广泛扩展的miRNA列表。有关保守和特定谱系的miRNA在蚊子中的进化获得了新的见解。一些miRNA的表达谱表明特定阶段的功能以及与胚胎发育或血液喂养相关的功能。更好地了解这些miRNA的功能将为蚊子生物学提供新的见解，并可能导致对抗蚊子传播的传染病的新方法。

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