Wang HaiYang, Xiao ShenHua, Wang Min, Kim Nam-Hyung, Li HuiXia, Wang GenLin
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China; Department of Animal Sciences, Chungbuk National University, Naesudong-ro, Seowon-gu, Cheongju-si 362-763, Chungcheongbuk-do, Republic of Korea.
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
Gene. 2015 Apr 1;559(2):119-28. doi: 10.1016/j.gene.2015.01.021. Epub 2015 Jan 24.
MicroRNAs (miRNAs) represent a newly identified class of non-protein-coding ~22 nt small RNA which plays important roles in multiple biological processes by degrading targeted mRNA or repressing mRNA translation. Here we present EST (expressed sequence tags)-based and GSS (Genomic Survey Sequences)-based combined approach for the detection of conserved miRNAs of cattle. A total of 20 conserved miRNAs that belong to 18 miRNA families were detected following a range of filtering criteria; their functions were further predicted and analyzed. To confirm our prediction, a miRNA-detecting microarray was designed with probes complementary to previously known mature miRNA sequences from 131 organisms. After hybridizing with small RNAs extracted from beef cattle subcutaneous fat tissue, 219 (32.30%) miRNAs were detected in the 679 known Bos taurus miRNAs and all the miRNAs predicted above were also detected. Conformation of 22 most abundant miRNA expression by qRT-PCR indicated that they were highly accumulated not only in subcutaneous fat tissue but also in intramuscular fat tissue. Bioinformatics of KEGG pathway analysis suggested that 4 differential expression miRNAs (miR-143, miR-145, miR-2325c and miR-2361) involved in different pathways and target genes may regulate the fat deposition differently. Taken together, our results expand the number of known bovine miRNAs and provide a thorough account of the miRNA transcriptome in bovine adipose tissue.
微小RNA（miRNA）是一类新发现的非蛋白质编码的约22个核苷酸的小RNA，通过降解靶mRNA或抑制mRNA翻译在多种生物学过程中发挥重要作用。在此，我们提出了基于EST（表达序列标签）和GSS（基因组调查序列）的联合方法来检测牛的保守miRNA。按照一系列筛选标准，共检测到属于18个miRNA家族的20个保守miRNA；对其功能进行了进一步预测和分析。为了证实我们的预测，设计了一种miRNA检测微阵列，其探针与来自131种生物的先前已知的成熟miRNA序列互补。与从肉牛皮下脂肪组织中提取的小RNA杂交后，在679个已知的牛miRNA中检测到219个（32.30%）miRNA，并且上述预测的所有miRNA也被检测到。通过qRT-PCR对22种最丰富的miRNA表达进行的验证表明，它们不仅在皮下脂肪组织中高度积累，而且在肌内脂肪组织中也高度积累。KEGG通路分析的生物信息学表明，4种差异表达的miRNA（miR-143、miR-145、miR-2325c和miR-2361）参与不同的通路，其靶基因可能对脂肪沉积有不同的调节作用。综上所述，我们的结果扩展了已知牛miRNA的数量，并全面描述了牛脂肪组织中的miRNA转录组。
