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Lars Dölken 1 , Zsolt Ruzsics 1 , Bernd Rädle 1 , Caroline C. Friedel 2 , Ralf Zimmer 2 , Jörg Mages 3 , Reinhard Hoffmann 3 , Paul Dickinson transient house in baguio 4 , Thorsten Forster 4 , Peter Ghazal 4 , and Ulrich H. Koszinowski 1 1 Max von Pettenkofer-Institute, Ludwig Maximilians-University Munich, Munich 80337, Germany 2 Institute for Informatics, Ludwig Maximilians-University Munich, Munich transient house in baguio 80333, Germany 3 Institute of Medical Microbiology, Technical University Munich, Munich 81675, Germany 4 Division of Pathway Medicine and Centre for Systems Biology at Edinburgh, The University transient house in baguio of Edinburgh, Edinburgh, Scotland EH16 4SB, United Kingdom transient house in baguio
RNA levels in a cell are determined by the relative rates of RNA synthesis and decay. State-of-the-art transcriptional analyses only employ total cellular RNA. Therefore, changes in RNA levels cannot be attributed to RNA synthesis or decay, and temporal resolution is poor. Recently, it was reported that newly transcribed RNA can be biosynthetically labeled transient house in baguio

for 1 2 h using thiolated nucleosides,

purified from total cellular RNA and subjected to microarray analysis. However, in order to study signaling events at molecular level, analysis of changes occurring within minutes is required. We developed an improved approach to separate total cellular RNA into newly transcribed and preexisting RNA following 10 15 min of metabolic labeling. Employing new computational tools for array normalization and half-life determination we simultaneously study short-term RNA synthesis and decay as well as their impact on cellular transcript levels. transient house in baguio As an example we studied the response of fibroblasts to type I and II interferons (IFN). Analysis of RNA transcribed within 15 30 min at different times during the first three hours of interferon-receptor activation resulted in a >10-fold increase in microarray sensitivity and provided a comprehensive profile of the kinetics of IFN-mediated changes in gene expression. We identify a previously undisclosed highly connected network of short-lived transcripts selectively down-regulated by IFNγ in between 30 and 60 min after IFN treatment showing transient house in baguio strong associations with cell cycle and apoptosis, indicating novel mechanisms by which IFNγ affects these pathways. Keywords microarray biosynthetic labeling half-life 4-thiouridine interferon
Current Issue
From the Cover AUF1/hnRNP D is a novel protein partner of the EBER1 noncoding RNA of Epstein-Barr virus Intrinsic flexibility of snRNA hairpin loops facilitates protein binding A comprehensive analysis of precursor microRNA cleavage by human Dicer TRBP alters human precursor microRNA processing rates and product length in vitro Cas5d processes pre-crRNA and is a member of a larger family of CRISPR RNA endonucleases