Department of Cell and Systems Biology/CAGEF, University of Toronto, Toronto, ON, Canada.
University of North Carolina at Charlotte, Charlotte, NC, USA.
Plant J. 2019 Nov;100(3):641-654. doi: 10.1111/tpj.14468. Epub 2019 Aug 23.
Improvements in next-generation sequencing technologies have resulted in dramatically reduced sequencing costs. This has led to an explosion of '-seq'-based methods, of which RNA sequencing (RNA-seq) for generating transcriptomic data is the most popular. By analysing global patterns of gene expression in organs/tissues/cells of interest or in response to chemical or environmental perturbations, researchers can better understand an organism's biology. Tools designed to work with large RNA-seq data sets enable analyses and visualizations to help generate hypotheses about a gene's function. We present here a user-friendly RNA-seq data exploration tool, called the 'eFP-Seq Browser', that shows the read map coverage of a gene of interest in each of the samples along with 'electronic fluorescent pictographic' (eFP) images that serve as visual representations of expression levels. The tool also summarizes the details of each RNA-seq experiment, providing links to archival databases and publications. It automatically computes the reads per kilobase per million reads mapped expression-level summaries and point biserial correlation scores to sort the samples based on a gene's expression level or by how dissimilar the read map profile is from a gene splice variant, to quickly identify samples with the strongest expression level or where alternative splicing might be occurring. Links to the Integrated Genome Browser desktop visualization tool allow researchers to visualize and explore the details of RNA-seq alignments summarized in eFP-Seq Browser as coverage graphs. We present four cases of use of the eFP-Seq Browser for ABI3, SR34, SR45a and U2AF65B, where we examine expression levels and identify alternative splicing. The URL for the browser is https://bar.utoronto.ca/eFP-Seq_Browser/. OPEN RESEARCH BADGES: This article has earned an Open Data Badge for making publicly available the digitally-shareable data necessary to reproduce the reported results. Tool is at https://bar.utoronto.ca/eFP-Seq_Browser/; RNA-seq data at https://s3.amazonaws.com/iplant-cdn/iplant/home/araport/rnaseq_bam/ and https://s3.amazonaws.com/iplant-cdn/iplant/home/araport/rnaseq_bam/Klepikova/. Code is available at https://github.com/BioAnalyticResource/eFP-Seq-Browser.
下一代测序技术的改进使得测序成本大幅降低。这导致了基于 '-seq' 的方法的爆炸式增长,其中 RNA 测序(RNA-seq)生成转录组数据是最受欢迎的。通过分析感兴趣的器官/组织/细胞中基因表达的全局模式,或对化学或环境扰动的反应,研究人员可以更好地了解生物体的生物学。专门用于处理大型 RNA-seq 数据集的工具可以进行分析和可视化,以帮助生成有关基因功能的假设。我们在这里介绍一个用户友好的 RNA-seq 数据探索工具,称为“eFP-Seq Browser”,它显示了每个样本中感兴趣基因的读取映射覆盖,并带有“电子荧光象形图”(eFP)图像,作为表达水平的视觉表示。该工具还总结了每个 RNA-seq 实验的详细信息,提供了对存档数据库和出版物的链接。它自动计算每百万映射读取的每千碱基的表达水平摘要和双位点相关系数,以便根据基因的表达水平或读取映射图谱与基因剪接变体的差异程度对样本进行排序,快速识别表达水平最强的样本或可能发生选择性剪接的样本。链接到集成基因组浏览器桌面可视化工具允许研究人员可视化和探索 eFP-Seq Browser 中总结的 RNA-seq 对齐的详细信息,作为覆盖图。我们提出了使用 eFP-Seq Browser 检查 ABI3、SR34、SR45a 和 U2AF65B 的四个案例,其中我们检查了表达水平并识别了选择性剪接。浏览器的 URL 是 https://bar.utoronto.ca/eFP-Seq_Browser/。开放研究徽章:本文为再现报告结果而公开了可重复数据共享的数字共享数据,因此获得了开放数据徽章。工具位于 https://bar.utoronto.ca/eFP-Seq_Browser/;RNA-seq 数据位于 https://s3.amazonaws.com/iplant-cdn/iplant/home/araport/rnaseq_bam/ 和 https://s3.amazonaws.com/iplant-cdn/iplant/home/araport/rnaseq_bam/Klepikova/。代码位于 https://github.com/BioAnalyticResource/eFP-Seq-Browser。