Zürich-Basel Plant Science Center, University of Basel, Department of Environmental Sciences, Botany, Hebelstrasse 1, 4056 Basel, Switzerland; Fasteris SA, Ch. du Pont-du-Centenaire 109, 1228 Plan-les-Ouates, Switzerland.
J Virol Methods. 2014 Jan;195:120-2. doi: 10.1016/j.jviromet.2013.10.013. Epub 2013 Oct 14.
In eukaryotes, diverse small RNA (sRNA) populations including miRNAs, siRNAs and piRNAs regulate gene expression and repress transposons, transgenes and viruses. Functional sRNAs are associated with effector proteins based on their size and nucleotide composition. The sRNA populations are currently analyzed by deep sequencing that generates millions of reads which are then mapped to a reference sequence or database. Here we developed a tool called MISIS to view and analyze sRNA maps of genomic loci and viruses which spawn multiple sRNAs. MISIS displays sRNA reads as a histogram where the x-axis indicates positions of the 5'- or 3'-terminal nucleotide of sense and antisense sRNAs, respectively, along a given reference sequence or its selected region and the y-axis the number of reads starting (for sense sRNA) or ending (for antisense sRNA) at each position. Size-classes of sRNAs can be visualized and compared separately or in combination. Thus, MISIS gives an overview of sRNA distribution along the reference sequence as well as detailed information on single sRNA species of different size-classes and abundances. MISIS reads standard BAM/SAM files outputted by mapping tools and generates table files containing counts of sRNA reads at each position of the reference sequence forward and reverse strand and for each of the chosen size-classes of sRNAs. These table files can be used by other tools such as Excel for further quantitative analysis and visualization. MISIS is a Java standalone program. It is freely available along with the source code at the following website: http://www.fasteris.com/apps.
在真核生物中,多种小 RNA(sRNA)群体,包括 miRNA、siRNA 和 piRNA,调节基因表达并抑制转座子、转座基因和病毒。功能 sRNA 与效应蛋白相关,基于它们的大小和核苷酸组成。sRNA 群体目前通过深度测序进行分析,该技术会生成数百万个读数,然后将这些读数映射到参考序列或数据库。在这里,我们开发了一种名为 MISIS 的工具,用于查看和分析基因组基因座和产生多种 sRNA 的病毒的 sRNA 图谱。MISIS 将 sRNA 读数显示为一个直方图,其中 x 轴表示给定参考序列或其选定区域中正义和反义 sRNA 的 5' 或 3' 末端核苷酸的位置,y 轴表示在每个位置开始(对于正义 sRNA)或结束(对于反义 sRNA)的读数数量。可以单独或组合可视化和比较 sRNA 的大小类别。因此,MISIS 提供了参考序列沿线 sRNA 分布的概述,以及关于不同大小类别和丰度的单个 sRNA 物种的详细信息。MISIS 读取映射工具输出的标准 BAM/SAM 文件,并生成包含参考序列正向和反向链上每个位置的 sRNA 读数计数以及所选 sRNA 大小类别的每个读数计数的表文件。这些表文件可以由其他工具(如 Excel)用于进一步的定量分析和可视化。MISIS 是一个 Java 独立程序。它可在以下网站上免费获得:http://www.fasteris.com/apps。