Gong Wangbao, Huang Yong, Xie Jun, Wang Guangjun, Yu Deguang, Sun Xihong
Key Laboratory of Tropical & Subtropical Fishery Resource Application & Cultivation, Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510380, China.
College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, 471003, China.
Comput Biol Chem. 2017 Jun;68:92-100. doi: 10.1016/j.compbiolchem.2017.02.010. Epub 2017 Mar 2.
MicroRNAs (miRNAs) are post-transcriptional regulators which bind to target to regulate protein expression by repressing translation or promoting degradation of the target mRNA. Studies have shown that deep sequencing is a powerful tool for the identification of miRNAs, and it is believed that may more miRNAs remain to be discovered in grass carp. In the present study, a pool of equal amounts of RNA obtained from 8 different adult grass carp tissues (spleen, liver, muscle, kidney, skin, testis, gut and heart) was sequenced using deep sequencing technology. A total of 16.579.334 raw reads were yielded from the pooled small RNA library. Using bioinformatics analysis, we identified 160 conserved miRNAs and 18 novel miRNAs in grass carp. Randomly selected 6 conserved and 2 novel miRNAs were confirmed their expression by stem-loop qRT-PCR assay. Furthermore, the 1212 potential targets of these miRNAs were predicted using miRNA target prediction tool. GO and KEGG pathway enrichment analyses indicated relevant biological processes. Our study provides the first genome-wide investigation of miRNAs from 8 mixed tissues of grass carp, and the data obtained expand the known grass carp miRNA inventory and provide a basis for further understanding functions of grass carp miRNAs.
微小RNA(miRNA)是转录后调节因子,其通过与靶标结合,抑制翻译或促进靶标mRNA降解来调节蛋白质表达。研究表明,深度测序是鉴定miRNA的有力工具,并且据信在草鱼中可能还有更多miRNA有待发现。在本研究中,使用深度测序技术对从8种不同成年草鱼组织(脾脏、肝脏、肌肉、肾脏、皮肤、睾丸、肠道和心脏)中获得的等量RNA池进行测序。从混合的小RNA文库中总共产生了16,579,334条原始读数。通过生物信息学分析,我们在草鱼中鉴定出160个保守miRNA和18个新miRNA。通过茎环定量逆转录聚合酶链反应(qRT-PCR)分析确认了随机选择的6个保守miRNA和2个新miRNA的表达。此外,使用miRNA靶标预测工具预测了这些miRNA的1212个潜在靶标。基因本体(GO)和京都基因与基因组百科全书(KEGG)通路富集分析表明了相关的生物学过程。我们的研究首次对草鱼8种混合组织中的miRNA进行了全基因组研究,所获得的数据扩展了已知的草鱼miRNA清单,并为进一步了解草鱼miRNA的功能提供了基础。