SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.

mRNA synthesis, processing, and destruction involve a complex series of molecular steps that are incompletely understood. Because the RNA intermediates in each of these steps have finite lifetimes, extensive mechanistic and dynamical information is encoded in total cellular RNA. Here we report the d...

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Main Authors: Jesse M Gray, David A Harmin, Sarah A Boswell, Nicole Cloonan, Thomas E Mullen, Joseph J Ling, Nimrod Miller, Scott Kuersten, Yong-Chao Ma, Steven A McCarroll, Sean M Grimmond, Michael Springer
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089673&type=printable
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author Jesse M Gray
David A Harmin
Sarah A Boswell
Nicole Cloonan
Thomas E Mullen
Joseph J Ling
Nimrod Miller
Scott Kuersten
Yong-Chao Ma
Steven A McCarroll
Sean M Grimmond
Michael Springer
author_facet Jesse M Gray
David A Harmin
Sarah A Boswell
Nicole Cloonan
Thomas E Mullen
Joseph J Ling
Nimrod Miller
Scott Kuersten
Yong-Chao Ma
Steven A McCarroll
Sean M Grimmond
Michael Springer
author_sort Jesse M Gray
collection DOAJ
description mRNA synthesis, processing, and destruction involve a complex series of molecular steps that are incompletely understood. Because the RNA intermediates in each of these steps have finite lifetimes, extensive mechanistic and dynamical information is encoded in total cellular RNA. Here we report the development of SnapShot-Seq, a set of computational methods that allow the determination of in vivo rates of pre-mRNA synthesis, splicing, intron degradation, and mRNA decay from a single RNA-Seq snapshot of total cellular RNA. SnapShot-Seq can detect in vivo changes in the rates of specific steps of splicing, and it provides genome-wide estimates of pre-mRNA synthesis rates comparable to those obtained via labeling of newly synthesized RNA. We used SnapShot-Seq to investigate the origins of the intrinsic bimodality of metazoan gene expression levels, and our results suggest that this bimodality is partly due to spillover of transcriptional activation from highly expressed genes to their poorly expressed neighbors. SnapShot-Seq dramatically expands the information obtainable from a standard RNA-Seq experiment.
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institution OA Journals
issn 1932-6203
language English
publishDate 2014-01-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS ONE
spelling doaj-art-1a3d01b1100d44b6a39d16e65f33d4812025-08-20T02:15:24ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0192e8967310.1371/journal.pone.0089673SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.Jesse M GrayDavid A HarminSarah A BoswellNicole CloonanThomas E MullenJoseph J LingNimrod MillerScott KuerstenYong-Chao MaSteven A McCarrollSean M GrimmondMichael SpringermRNA synthesis, processing, and destruction involve a complex series of molecular steps that are incompletely understood. Because the RNA intermediates in each of these steps have finite lifetimes, extensive mechanistic and dynamical information is encoded in total cellular RNA. Here we report the development of SnapShot-Seq, a set of computational methods that allow the determination of in vivo rates of pre-mRNA synthesis, splicing, intron degradation, and mRNA decay from a single RNA-Seq snapshot of total cellular RNA. SnapShot-Seq can detect in vivo changes in the rates of specific steps of splicing, and it provides genome-wide estimates of pre-mRNA synthesis rates comparable to those obtained via labeling of newly synthesized RNA. We used SnapShot-Seq to investigate the origins of the intrinsic bimodality of metazoan gene expression levels, and our results suggest that this bimodality is partly due to spillover of transcriptional activation from highly expressed genes to their poorly expressed neighbors. SnapShot-Seq dramatically expands the information obtainable from a standard RNA-Seq experiment.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089673&type=printable
spellingShingle Jesse M Gray
David A Harmin
Sarah A Boswell
Nicole Cloonan
Thomas E Mullen
Joseph J Ling
Nimrod Miller
Scott Kuersten
Yong-Chao Ma
Steven A McCarroll
Sean M Grimmond
Michael Springer
SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.
PLoS ONE
title SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.
title_full SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.
title_fullStr SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.
title_full_unstemmed SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.
title_short SnapShot-Seq: a method for extracting genome-wide, in vivo mRNA dynamics from a single total RNA sample.
title_sort snapshot seq a method for extracting genome wide in vivo mrna dynamics from a single total rna sample
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0089673&type=printable
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