使用poly(A)-tailed RT-PCR对猪部分microRNAs在胚胎期的表达进行验证

Identification Pig Partial MicroRNAs at Embryo by Poly(A)-tailed RT-PCR

MicroRNAs(miRNAs)是一系列小的非编码的RNA,长度大约为21～22nt。主要通过抑制翻译或者进行mRNA的降解而在多种生理以及发育过程中发挥很重要的作用。miRB ase16.0(2010)数据库中显示,注释的线虫、小鼠、大鼠以及人的miRNAs分别有175种、673种、408种和1048种。而哺乳动物猪的miRNAs仅仅为211种,特别是在胚胎生长发育过程中调节基因表达的miRNAs的研究十分欠缺。因此使用poly(A)-tailed RT-PCR对猪的10种miRNAs的表达在猪胚胎期第33天(E33)进行验证。对于鉴定的这10种miRNAs,通过实时定量PCR对其表达水平进行相对分析,并使用PicTar和TargetScan软件对其靶基因进行预测。结果表明:在头部组织中ssc-miR-9的表达量最高的,而在内脏组织中ssc-miR-1的表达量最高。功能分析表明这些miRNAs可以参与神经、大脑、肌肉、器官的发育。此研究为以后进一步的研究猪以及相关动物胚胎发育的miRNAs的功能提供有用的信息。

MicroRNAs （miRNAs） are a class of small noncoding RNAs of 21~22 nucleotides in length that play important roles in multiple biological and developmental processes by degrading targeted mRNAs or repressing mRNA translation. The database of miRBase16.0 （2010） show that there are 175, 673, 408, and 1048 annotated miRNAs for Caenorhabditis elegans, Mus musculus, Rattus norvegicus, and Homo sapiens, respectively. However, there are only 211 miRNAs described for Sus scrofa. In particular, the miRNAs which regulate gene expression in embryonic growth and development were still poorly understood. Therefore, in this study we carried out poly（A）-tailed RT-PCR to verify the expression of ten miRNAs at porcine embryonic day 33 （E33）. Of these distinct miRNAs identified, the author determined the relative amount of miRNAs by quantitative real-time PCR and performed investigation into identification of the potential target mRNAs using PicTar and TargetScan. Results showed that： ssc-miR-9 was the most abundantly expressed miRNA in the head region, ssc-miR-1 was the most abundantly expressed miRNA in the organ region. Function analysis suggested thatthese miRNAs were mostly involved in the development of nerves, the cerebrum, muscle, and organs. The results provide useful information for the investigation of embryonic miRNAs in the pig and other animals.