Chapter title |
Enhancer RNAs
|
---|---|
Chapter number | 9 |
Book title |
Enhancer RNAs
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-4035-6_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-4033-2, 978-1-4939-4035-6
|
Authors |
Gardini, Alessandro, Alessandro Gardini Ph.D., Alessandro Gardini |
Editors |
Ulf Andersson Ørom |
Abstract |
Transcription occurring at gene loci results in accumulation of mature RNA molecules (i.e., mRNAs) that can be easily assayed by RT-PCR or RNA sequencing. However, the steady-state level of RNA does not accurately mirror transcriptional activity per se. In fact, RNA stability plays a major role in determining the relative abundance of any given RNA molecule. Here, I describe a protocol of Nuclear Run-On assay coupled to deep sequencing to assess real-time transcription from engaged RNA polymerase. Mapping nascent transcripts at the genome-wide scale provides a reliable measure of transcriptional activity in mammalian cells and delivers a high-resolution map of coding and noncoding transcripts that is especially useful for annotation and quantification of short-lived RNA molecules. |
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