TAGC, Case 928, 163 Avenue de Luminy, 13288 Marseille Cedex 09, France; Aix-Marseille Université, Marseille, France; Inserm, UMR1090, Marseille, France.
Methods. 2013 Sep 1;63(1):3-17. doi: 10.1016/j.ymeth.2013.04.003. Epub 2013 Apr 15.
The explosion of high throughput sequencing technologies marked a turn in our way of understanding the complexity and diversity of the transcriptome, including noncoding transcription dependent on RNA polymerase II. Many new ncRNA populations were described in recent years, including for example TSS RNAs, lincRNAs, eRNAs, PROMPTS and several others. Besides the advances in the average depth coverage of RNA-seq experiments, various additional protocols are now available that can be used to address qualitative and quantitative aspects of the noncoding transcriptome complexity and function. In this review, we will focus on methods allowing isolation and characterization of complex RNA populations using sequencing based approaches, including conventional strategies already used for coding genome and more specific developments allowing, for example, the study of nascent strand transcription, protein-bound or structured RNAs.
高通量测序技术的爆炸式发展标志着我们理解转录组复杂性和多样性方式的转变,包括依赖 RNA 聚合酶 II 的非编码转录。近年来描述了许多新的 ncRNA 群体,例如 TSS RNA、lincRNA、eRNA、PROMPTS 等。除了 RNA-seq 实验平均深度覆盖的提高外,现在还提供了各种其他的方案,可以用于解决非编码转录组复杂性和功能的定性和定量方面。在这篇综述中,我们将重点介绍使用基于测序的方法分离和表征复杂 RNA 群体的方法,包括已经用于编码基因组的常规策略和更具体的发展,例如,研究新生链转录、蛋白质结合或结构 RNA。