State Key Laboratory of Agrobiotechnology, Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
State Key Laboratory of Agrobiotechnology, Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
Gene. 2014 Apr 15;539(2):224-9. doi: 10.1016/j.gene.2014.01.074. Epub 2014 Feb 11.
MicroRNAs (miRNAs) are small regulatory RNAs that play a significant role in eukaryotes by targeting mRNAs for cleavage or translational repression. Recent studies have also shown them to be associated with cellular changes following viral infection. Mink enteritis virus (MEV) is one of the most important viral pathogens in the mink industry. To study the involvement of miRNAs in the MEV infection process, we used Illumina's ultrahigh throughput approach to sequencing miRNA libraries from the feline kidney (F81) cell line before and after infection with MEV. Using this bioinformatics approach we identified 196 known mammalian miRNA orthologs belonging to 152 miRNA families in F81 cells. Additionally, 97 miRNA*s of these miRNAs were detected. As well as known miRNAs, 384 and 398 novel miRNA precursor candidates were identified in uninfected and MEV-infected F81 cells respectively that have not been reported in other mammals. In MEV-infected cells 3 miRNAs were significantly down-regulated and 4 up-regulated including 3 significantly. The majority (12 of 16) of randomly selected miRNA expression profiles by qRT-PCR were consistent with those identified by deep sequencing. A total of 88 miRNAs were predicted to target interferon-associated genes; 6 appear to target the 3'UTR of MEV-specific receptor transferring receptor mRNAs; and 8 to target the MEV mRNA coding region. No miRNAs coded by MEV itself were detected.
微小 RNA（miRNAs）是一类在真核生物中起重要作用的小调控 RNA，通过靶向 mRNAs 进行切割或翻译抑制来发挥作用。最近的研究还表明，它们与病毒感染后细胞的变化有关。水貂肠炎病毒（MEV）是水貂产业中最重要的病毒病原体之一。为了研究 miRNAs 在 MEV 感染过程中的作用，我们使用 Illumina 的超高通量方法，对感染 MEV 前后的猫肾（F81）细胞系的 miRNA 文库进行测序。使用这种生物信息学方法，我们在 F81 细胞中鉴定出 196 个已知的哺乳动物 miRNA 直系同源物，属于 152 个 miRNA 家族。此外，还检测到这些 miRNA 的 97 个 miRNA*。除了已知的 miRNAs，在未感染和 MEV 感染的 F81 细胞中分别鉴定出 384 和 398 个新的 miRNA 前体候选物，这些候选物在其他哺乳动物中尚未报道过。在 MEV 感染的细胞中，有 3 个 miRNA 显著下调，4 个 miRNA 上调，其中 3 个显著上调。通过 qRT-PCR 随机选择的 16 个 miRNA 表达谱中的 12 个与深度测序鉴定的结果一致。共有 88 个 miRNA 被预测靶向干扰素相关基因；6 个似乎靶向 MEV 特异性受体转移受体 mRNAs 的 3'UTR；8 个靶向 MEV mRNA 编码区。没有检测到 MEV 自身编码的 miRNAs。
