不同体长淡水虾微小RNA的特征及其差异表达

Pub Date : 2020-01-02 DOI:10.1080/07924259.2019.1694085
Ran Li, Xin Wang, Qinghao Meng, Liqi Ren, Jieyang Weng, Shen Wang, Jinsheng Sun
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引用次数: 2

摘要

MicroRNAs (miRNAs)是一类促进mRNA降解和抑制mRNA翻译的小非编码rna。通过转录后调控,microRNAs (miRNAs)在调节生长、发育、代谢和生殖过程中发挥重要作用。然而,目前对甲壳类动物中mirna的了解有限。在这项研究中,我们探讨了mirna在不同体长的异足新蜥腹肌中的潜在作用。我们从肌肉样本中构建了两个小RNA文库,并鉴定了26个新的mirna。在这些新的mirna中,有13个被发现有显著差异表达。其中,10种mirna在小虾中显著上调,3种mirna在大虾中显著下调。此外,生物信息学分析显示miR-26与大量差异表达基因相关,包括葡萄糖转运蛋白1 (Glut1)、磷酸烯醇丙酮酸羧激酶(Pepck)和磷酸果糖激酶(Pfk),这些基因与糖酵解和糖异生途径相关。此外,荧光素酶活性测定证实了Pepck是新型miR-26的直接靶基因。研究结果提示miRNA具有潜在的生长调控作用,为进一步研究异足奈米和其他甲壳类动物肌肉生长的miRNA调控机制提供了宝贵的资源。缩写:Ago; argonaute;ANOVA:单因素方差分析;CRISPR:聚集规则间隔的短回文重复序列;DEGs:差异表达基因;DMEM: Dulbecco 's Modified Eagle Medium;Glut1:葡萄糖转运蛋白1;GO:基因本体;京都基因与基因组百科全书;microRNA:微;磷酸烯醇丙酮酸羧激酶;ppi:预测蛋白-蛋白相互作用;Pfk:磷酸果糖激酶;RISC: rna诱导沉默复合体;sirna:小干扰rna;sRNA:小RNA;dsDNA:双链DNA;qPCR:定量PCR
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Characterization and differential expression of microRNA in the freshwater shrimp (Neocaridina heteropoda) with different body length
ABSTRACT MicroRNAs (miRNAs) are a class of small non-coding RNAs which promote mRNA degradation and inhibit mRNA translation. Through post-transcriptional regulation, microRNAs (miRNAs) play a significant role in regulating growth, development, metabolic, and reproductive processes. However, current knowledge on miRNAs in crustaceans is limited. In this study, we explored the potential roles of miRNAs in abdominal muscle of Neocaridina heteropoda of different body length. We constructed two small RNA libraries from muscle samples, and 26 novel miRNAs were identified. Among these novel miRNAs, 13 were found significantly differentially expressed. Specifically, 10 miRNAs were significantly up-regulated and 3 miRNAs were down-regulated in small shrimps compared with large ones. Furthermore, bioinformatics analysis revealed that miR-26 was associated with a large number of differentially expressed genes, including glucose transporter 1 (Glut1), phosphoenolpyruvate carboxykinase (Pepck), and phosphofructokinase (Pfk), related to glycolysis and gluconeogenesis pathways. Besides, luciferase activity assay verified that Pepck was a direct target gene of the novel miR-26. The results imply potential growth regulatory roles for miRNAs and provide a valuable resource for further investigation of the miRNA mechanisms controlling muscle growth in N. heteropoda and other crustaceans. Abbreviations: Ago: argonaute; ANOVA: one-way analysis of variance; CRISPR: clustered regularly interspaced short palindromic repeats; DEGs: differentially expressed genes; DMEM: Dulbecco’s Modified Eagle Medium; Glut1: glucose transporter 1; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; miRNA: microRNA; Pepck: phosphoenolpyruvate carboxykinase; PPIs: predict protein-protein interactions; Pfk: phosphofructokinase; RISC: RNA-induced silencing complex; siRNAs: small interfering RNAs; sRNA: small RNA; dsDNA: double-stranded DNA; qPCR: quantitative PCR
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