Division of Biological Sciences, University of California San Diego, La Jolla, California 92093, USA.
RNA. 2022 May;28(5):645-656. doi: 10.1261/rna.079071.121. Epub 2022 Feb 18.
Post-transcriptional trimming and tailing of RNA 3' ends play key roles in the processing and quality control of noncoding RNAs (ncRNAs). However, bioinformatic tools to examine changes in the RNA 3' "tailome" are sparse and not standardized. Here we present Tailer, a bioinformatic pipeline in two parts that allows for robust quantification and analysis of tail information from next-generation sequencing experiments that preserve RNA 3' end information. The first part of Tailer, Tailer-processing, uses genome annotation or reference FASTA gene sequences to quantify RNA 3' ends from SAM-formatted alignment files or FASTQ sequence read files produced from sequencing experiments. The second part, Tailer-analysis, uses the output of Tailer-processing to identify statistically significant RNA targets of trimming and tailing and create graphs for data exploration. We apply Tailer to RNA 3' end sequencing experiments from three published studies and find that it accurately and reproducibly recapitulates key findings. Thus, Tailer should be a useful and easily accessible tool to globally investigate tailing dynamics of nonpolyadenylated RNAs and conditions that perturb them.
RNA 3' 端的转录后修剪和加尾在非编码 RNA(ncRNA)的加工和质量控制中起着关键作用。然而,用于检查 RNA 3'“尾组学”变化的生物信息学工具稀缺且未标准化。在这里,我们介绍了 Tailer,这是一个由两部分组成的生物信息学管道,允许从保留 RNA 3' 端信息的下一代测序实验中对尾信息进行稳健的定量和分析。Tailer 的第一部分 Tailer-processing 使用基因组注释或参考 FASTA 基因序列,从 SAM 格式的比对文件或测序实验产生的 FASTQ 序列读取文件中定量 RNA 3' 端。第二部分 Tailer-analysis 使用 Tailer-processing 的输出来识别修剪和加尾的具有统计学意义的 RNA 靶标,并创建图形以进行数据探索。我们将 Tailer 应用于来自三个已发表研究的 RNA 3' 端测序实验,发现它准确且可重复地再现了关键发现。因此,Tailer 应该是一种有用且易于访问的工具,可用于全面研究非多聚腺苷酸化 RNA 的加尾动态和扰乱它们的条件。