Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.

Pluripotent stem cells retain the developmental timing of their species of origin in vitro, an observation that suggests the existence of a cell-intrinsic developmental clock, yet the nature and machinery of the clock remain a mystery. We hypothesize that one possible component may lie in species-sp...

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Main Authors: Christopher Barry, Matthew T Schmitz, Cara Argus, Jennifer M Bolin, Mitchell D Probasco, Ning Leng, Bret M Duffin, John Steill, Scott Swanson, Brian E McIntosh, Ron Stewart, Christina Kendziorski, James A Thomson, Rhonda Bacher
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2019-12-01
Series:PLoS Computational Biology
Online Access:https://journals.plos.org/ploscompbiol/article/file?id=10.1371/journal.pcbi.1007543&type=printable
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author Christopher Barry
Matthew T Schmitz
Cara Argus
Jennifer M Bolin
Mitchell D Probasco
Ning Leng
Bret M Duffin
John Steill
Scott Swanson
Brian E McIntosh
Ron Stewart
Christina Kendziorski
James A Thomson
Rhonda Bacher
author_facet Christopher Barry
Matthew T Schmitz
Cara Argus
Jennifer M Bolin
Mitchell D Probasco
Ning Leng
Bret M Duffin
John Steill
Scott Swanson
Brian E McIntosh
Ron Stewart
Christina Kendziorski
James A Thomson
Rhonda Bacher
author_sort Christopher Barry
collection DOAJ
description Pluripotent stem cells retain the developmental timing of their species of origin in vitro, an observation that suggests the existence of a cell-intrinsic developmental clock, yet the nature and machinery of the clock remain a mystery. We hypothesize that one possible component may lie in species-specific differences in the kinetics of transcriptional responses to differentiation signals. Using a liquid-handling robot, mouse and human pluripotent stem cells were exposed to identical neural differentiation conditions and sampled for RNA-sequencing at high frequency, every 4 or 10 minutes, for the first 10 hours of differentiation to test for differences in transcriptomic response rates. The majority of initial transcriptional responses occurred within a rapid window in the first minutes of differentiation for both human and mouse stem cells. Despite similarly early onsets of gene expression changes, we observed shortened and condensed gene expression patterns in mouse pluripotent stem cells compared to protracted trends in human pluripotent stem cells. Moreover, the speed at which individual genes were upregulated, as measured by the slopes of gene expression changes over time, was significantly faster in mouse compared to human cells. These results suggest that downstream transcriptomic response kinetics to signaling cues are faster in mouse versus human cells, and may offer a partial account for the vast differences in developmental rates across species.
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issn 1553-734X
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spelling doaj-art-14bcc2d6b0e84205bbe5dfa387cbcb922025-08-20T02:45:50ZengPublic Library of Science (PLoS)PLoS Computational Biology1553-734X1553-73582019-12-011512e100754310.1371/journal.pcbi.1007543Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.Christopher BarryMatthew T SchmitzCara ArgusJennifer M BolinMitchell D ProbascoNing LengBret M DuffinJohn SteillScott SwansonBrian E McIntoshRon StewartChristina KendziorskiJames A ThomsonRhonda BacherPluripotent stem cells retain the developmental timing of their species of origin in vitro, an observation that suggests the existence of a cell-intrinsic developmental clock, yet the nature and machinery of the clock remain a mystery. We hypothesize that one possible component may lie in species-specific differences in the kinetics of transcriptional responses to differentiation signals. Using a liquid-handling robot, mouse and human pluripotent stem cells were exposed to identical neural differentiation conditions and sampled for RNA-sequencing at high frequency, every 4 or 10 minutes, for the first 10 hours of differentiation to test for differences in transcriptomic response rates. The majority of initial transcriptional responses occurred within a rapid window in the first minutes of differentiation for both human and mouse stem cells. Despite similarly early onsets of gene expression changes, we observed shortened and condensed gene expression patterns in mouse pluripotent stem cells compared to protracted trends in human pluripotent stem cells. Moreover, the speed at which individual genes were upregulated, as measured by the slopes of gene expression changes over time, was significantly faster in mouse compared to human cells. These results suggest that downstream transcriptomic response kinetics to signaling cues are faster in mouse versus human cells, and may offer a partial account for the vast differences in developmental rates across species.https://journals.plos.org/ploscompbiol/article/file?id=10.1371/journal.pcbi.1007543&type=printable
spellingShingle Christopher Barry
Matthew T Schmitz
Cara Argus
Jennifer M Bolin
Mitchell D Probasco
Ning Leng
Bret M Duffin
John Steill
Scott Swanson
Brian E McIntosh
Ron Stewart
Christina Kendziorski
James A Thomson
Rhonda Bacher
Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.
PLoS Computational Biology
title Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.
title_full Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.
title_fullStr Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.
title_full_unstemmed Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.
title_short Automated minute scale RNA-seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics.
title_sort automated minute scale rna seq of pluripotent stem cell differentiation reveals early divergence of human and mouse gene expression kinetics
url https://journals.plos.org/ploscompbiol/article/file?id=10.1371/journal.pcbi.1007543&type=printable
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