利用4-硫尿苷对新转录RNA进行代谢标记，对RNA合成、加工和衰变进行高分辨率基因表达谱分析。
High-Resolution Gene Expression Profiling of RNA Synthesis, Processing, and Decay by Metabolic Labeling of Newly Transcribed RNA Using 4-Thiouridine.
作者信息
Rutkowski Andrzej J, Dölken Lars
机构信息
Department of Medicine, University of Cambridge, Box 157, Level 5, Addenbrooke's Hospital, Hills Road, CB20QQ, Cambridge, UK.
Institut für Virologie und Immunbiologie, Julius-Maximilians-Universität Würzburg, Versbacher Str. 7, 97078, Würzburg, Germany.
出版信息
Methods Mol Biol. 2017;1507:129-140. doi: 10.1007/978-1-4939-6518-2_10.
Cellular RNA levels are orchestrated by highly regulated processes involving RNA synthesis (transcription), processing (e.g., splicing, polyadenylation, transport), and degradation. Profiling these changes provides valuable information on the regulation of gene expression. Total cellular RNA is a poor template for revealing short-term changes in gene expression, alterations in RNA decay rates, and the kinetics of RNA processing as well as the differentiation thereof. Here, we describe the metabolic labeling and purification of newly transcribed RNA with 4-thiouridine, by which these limitations are overcome.
细胞RNA水平由涉及RNA合成（转录）、加工（如剪接、聚腺苷酸化、转运）和降解的高度调控过程所协调。分析这些变化可为基因表达调控提供有价值的信息。总细胞RNA对于揭示基因表达的短期变化、RNA降解速率的改变、RNA加工动力学及其分化而言，是一种较差的模板。在此，我们描述了用4-硫尿苷对新转录的RNA进行代谢标记和纯化的方法，借此克服了这些局限性。
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