Characterisation of the small RNAs in the biomedically important green-bottle blowfly Lucilia sericata.
Characterisation of the small RNAs in the biomedically important green-bottle blowfly Lucilia sericata.
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DOI：
10.1371/journal.pone.0122203
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Phillips AR
中科院分区：
文献类型：
--
作者：
Blenkiron C;Tsai P;Brown LA;Tintinger V;Askelund KJ;Windsor JA;Phillips AR
丝光绿色蝇（Lucilia sericata）是一种重要的医学、农业和法医学物种。对这一物种生物学的进一步了解对许多研究团体都有很大的潜在好处。MicroRNA（miRNA）是一种短的非蛋白质编码调控RNA，在翻译水平上直接调控许多蛋白质编码基因。它们已被证明具有发育和组织特异性分布，它们直接影响基因调控。为了加深对L.我们已经对其不同发育阶段的分泌物和组织进行了小RNA测序。我们成功地从L.绢蚕蛆这些分泌物和三种组织（作物，唾液腺，肠道）的Illumina小RNA测序显示，最常见的小RNA片段来自核糖体RNA和转移RNA的昆虫和细菌的起源。这些RNA片段是高度特异性的，其中最常见的tRNA，如GlyGCC，主要由来自成熟蛆tRNA的5'端的读段代表。每个文库还具有独特的miRNA谱，其中在蛆分泌物和肠道中具有高丰度的miR-10- 5 p，并且在食物储存器官作物和唾液腺中具有高丰度的miR-8。生物活性蛆分泌物中的小RNA模式表明它们起源于唾液、前肠和后肠组织的组合。对RNA测序数据的液滴数字RT-PCR验证表明，不仅miRNA和小RNA片段的组织特征存在差异，而且这些差异也通过昆虫的发育阶段进行调节。我们鉴定了药用蛆L.研究表明，在特定组织中表达的miRNA有不同的子集，这些子集在昆虫发育过程中也会发生变化。此外，存在来自组织和分泌物中存在的其他非编码RNA的非常特异的RNA片段。这些新知识适用于不同的研究领域，包括伤口愈合，农业和法医学。
The green bottle fly maggot, Lucilia sericata, is a species with importance in medicine, agriculture and forensics. Improved understanding of this species’ biology is of great potential benefit to many research communities. MicroRNAs (miRNA) are a short non-protein coding regulatory RNA, which directly regulate a host of protein coding genes at the translational level. They have been shown to have developmental and tissue specific distributions where they impact directly on gene regulation. In order to improve understanding of the biology of L. sericata maggots we have performed small RNA-sequencing of their secretions and tissue at different developmental stages. We have successfully isolated RNA from the secretions of L. sericata maggots. Illumina small RNA-sequencing of these secretions and the three tissues (crop, salivary gland, gut) revealed that the most common small RNA fragments were derived from ribosomal RNA and transfer RNAs of both insect and bacterial origins. These RNA fragments were highly specific, with the most common tRNAs, such as GlyGCC, predominantly represented by reads derived from the 5’ end of the mature maggot tRNA. Each library also had a unique profile of miRNAs with a high abundance of miR-10-5p in the maggot secretions and gut and miR-8 in the food storage organ the crop and salivary glands. The pattern of small RNAs in the bioactive maggot secretions suggests they originate from a combination of saliva, foregut and hindgut tissues. Droplet digital RT-PCR validation of the RNA-sequencing data shows that not only are there differences in the tissue profiles for miRNAs and small RNA fragments but that these are also modulated through developmental stages of the insect. We have identified the small-RNAome of the medicinal maggots L. sericata and shown that there are distinct subsets of miRNAs expressed in specific tissues that also alter during the development of the insect. Furthermore there are very specific RNA fragments derived from other non-coding RNAs present in tissues and in the secretions. This new knowledge has applicability in diverse research fields including wound healing, agriculture and forensics.
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