Sun Wei, Chen Wei
Department of Biology, Southern University of Science and Technology, Shenzhen City, Nanshan District, China.
Laboratory for Functional and Medical Genomics, Berlin Institute for Medical Systems Biology, Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
Methods Mol Biol. 2018;1720:25-34. doi: 10.1007/978-1-4939-7540-2_3.
The development of genome-wide RNA profiling technologies greatly facilitates the global analysis of gene expression. However, such technologies alone could not distinguish the contribution to cellular RNA abundance by transcription versus decay. To overcome such limitation, metabolic labeling of newly synthesized RNA with 4-thiouridine (4sU) combined with genome-wide RNA profiling was used to in parallel measure RNA transcription and decay. Here, we describe the detailed protocol for using metabolic labeling with 4sU to separate newly synthesized RNA from the preexisting RNA in mammalian cells.
全基因组RNA分析技术的发展极大地促进了基因表达的全局分析。然而，仅靠这些技术无法区分转录和衰变对细胞RNA丰度的贡献。为了克服这一局限性，将新合成RNA用4-硫尿苷（4sU）进行代谢标记并结合全基因组RNA分析，用于并行测量RNA转录和衰变。在此，我们描述了使用4sU代谢标记从哺乳动物细胞中已有的RNA中分离新合成RNA的详细方案。
