High-Resolution Gene Expression Profiling of RNA Synthesis, Processing, and Decay by Metabolic Labeling of Newly Transcribed RNA Using 4-Thiouridine.
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.
Department of Medicine, University of Cambridge, Box 157, Level 5, Addenbrooke's Hospital, Hills Road, CB20QQ, Cambridge, UK.
Andrzej J
AJ
Rutkowski
Department of Medicine, University of Cambridge, Box 157, Level 5, Addenbrooke's Hospital, Hills Road, CB20QQ, Cambridge, UK. lars.doelken@uni-wuerzburg.de.|Institut für Virologie und Immunbiologie, Julius-Maximilians-Universität Würzburg, Versbacher Str. 7, 97078, Würzburg, Germany. lars.doelken@uni-wuerzburg.de.
Lars
L
Dölken
D000345:Affinity Labels / Q000302:isolation & purification / Q000378:metabolism; D002478:Cells, Cultured; D020869:Gene Expression Profiling / Q000379:methods*; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D013194:Staining and Labeling; D013891:Thiouridine / Q000302:isolation & purification* / Q000378:metabolism; D059467:Transcriptome
D000345:Affinity Labels; D012333:RNA, Messenger; D013891:Thiouridine
4-Thiouridine; Biotin; Gene expression profiling; Microarray; Newly transcribed RNA; RNA decay; RNA processing; RNA-seq; Streptavidin