Somatic piRNAs in the adult mouse

Small RNAs were deep sequenced from the liver and spleen of adult mice in an effort to identify somatic piRNAs. Following sequencing of all small RNAs, known non-coding RNAs were computationally removed from the dataset. The remaining RNAs were then mapped to the genome and analyzed for sequence characteristics (5' base, length) typical of known piRNAs. To determine if any of the identified small RNAs were MIWI2 dependent, we deep sequenced small RNAs from liver and spleen of MIWI2 KO mice and analyzed them as above. We deep sequenced small RNAs from the liver and spleen of one WT mouse and one MIWI2 knock-out mouse. We then trimmed sequencing adapters and removed known ncRNAs (rRNA, tRNA, snoRNA, snRNA, miRNA) from the dataset before aligning reads to the mm9 assembly of the mouse genome.为鉴定体细胞piRNA（Piwi-interacting RNA），本研究对成年小鼠的肝脏与脾脏组织中的小分子RNA进行了深度测序。完成全部小分子RNA测序后，通过计算方法移除了数据集中已知的非编码RNA（ncRNA）。随后将剩余RNA比对至参考基因组，并针对已知piRNA典型的序列特征（5'端碱基、序列长度）开展分析。为明确所鉴定的小分子RNA是否依赖MIWI2，本研究对MIWI2敲除（knockout, KO）小鼠的肝脏与脾脏组织中的小分子RNA进行深度测序，并按照前述分析流程开展分析。本研究同时对1只野生型（wild type, WT）小鼠与1只MIWI2敲除小鼠的肝脏、脾脏组织中的小分子RNA进行了深度测序。随后对测序数据进行接头修剪，并移除数据集中已知的非编码RNA（涵盖核糖体RNA（rRNA）、转运RNA（tRNA）、小核仁RNA（snoRNA）、小核RNA（snRNA）、微小RNA（miRNA）），再将测序读段（reads）比对至小鼠基因组mm9组装版本。