Gaining insight into transcriptome‐wide RNA population dynamics through the chemistry of 4‐thiouridine. (28th October 2018)

• Record Type: Journal Article
• Authors: Duffy, Erin E.; Schofield, Jeremy A.; Simon, Matthew D.
• Is Part Of: Wiley interdisciplinary reviews. Volume 10:Number 1(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-10-28
• Subjects: nucleoside recoding -- RNA metabolism -- RNA turnover -- transcriptome dynamics -- transient transcription; RNA -- Periodicals; 572.8805
• DOI: 10.1002/wrna.1513
• Languages: English
• ISSNs: 1757-7004

Abstract

Abstract : Cellular RNA levels are the result of a juggling act between RNA transcription, processing, and degradation. By tuning one or more of these parameters, cells can rapidly alter the available pool of transcripts in response to stimuli. While RNA sequencing (RNA‐seq) is a vital method to quantify RNA levels genome‐wide, it is unable to capture the dynamics of different RNA populations at steady‐state or distinguish between different mechanisms that induce changes to the steady‐state (i.e., altered rate of transcription vs. degradation). The dynamics of different RNA populations can be studied by targeted incorporation of noncanonical nucleosides. 4‐Thiouridine (s 4 U) is a commonly used and versatile RNA metabolic label that allows the study of many properties of RNA metabolism from synthesis to degradation. Numerous experimental strategies have been developed that leverage the power of s 4 U to label newly transcribed RNA in whole cells, followed by enrichment with activated disulfides or chemistry to induce C mutations at sites of s 4 U during sequencing. This review presents existing methods to study RNA population dynamics genome‐wide using s 4 U metabolic labeling, as well as a discussion of considerations and challenges when designing s 4 U metabolic labeling experiments. This article is categorized under: RNA Methods > RNA Analyses in Cells RNA Turnover and Surveillance > Regulation of RNA Stability Abstract : Newly transcribed RNAs can be metabolically labeled with 4‐thiouridine (s 4 U) to study many aspects of RNA metabolism genome‐wide, including RNA turnover, transient transcription, and polymerase elongation. New RNAs can be detected via biochemical enrichment (top) or nucleoside recoding to induce U‐to‐C mutations in s 4 U‐RNA in high‐throughput sequencing.