Rapid neurogenesis through transcriptional activation in human stem cells

Abstract Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yield...

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Main Authors: Volker Busskamp, Nathan E Lewis, Patrick Guye, Alex HM Ng, Seth L Shipman, Susan M Byrne, Neville E Sanjana, Jernej Murn, Yinqing Li, Shangzhong Li, Michael Stadler, Ron Weiss, George M Church
Format: Article
Language:English
Published: Springer Nature 2014-11-01
Series:Molecular Systems Biology
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Online Access:https://doi.org/10.15252/msb.20145508
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author Volker Busskamp
Nathan E Lewis
Patrick Guye
Alex HM Ng
Seth L Shipman
Susan M Byrne
Neville E Sanjana
Jernej Murn
Yinqing Li
Shangzhong Li
Michael Stadler
Ron Weiss
George M Church
author_facet Volker Busskamp
Nathan E Lewis
Patrick Guye
Alex HM Ng
Seth L Shipman
Susan M Byrne
Neville E Sanjana
Jernej Murn
Yinqing Li
Shangzhong Li
Michael Stadler
Ron Weiss
George M Church
author_sort Volker Busskamp
collection DOAJ
description Abstract Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yields. Here, we overexpressed two Neurogenin transcription factors in human‐induced pluripotent stem cells and obtained neurons with bipolar morphology in 4 days, at greater than 90% purity. The high purity enabled mRNA and microRNA expression profiling during neurogenesis, thus revealing the genetic programs involved in the rapid transition from stem cell to neuron. The resulting cells exhibited transcriptional, morphological and functional signatures of differentiated neurons, with greatest transcriptional similarity to prenatal human brain samples. Our analysis revealed a network of key transcription factors and microRNAs that promoted loss of pluripotency and rapid neurogenesis via progenitor states. Perturbations of key transcription factors affected homogeneity and phenotypic properties of the resulting neurons, suggesting that a systems‐level view of the molecular biology of differentiation may guide subsequent manipulation of human stem cells to rapidly obtain diverse neuronal types.
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institution Kabale University
issn 1744-4292
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publishDate 2014-11-01
publisher Springer Nature
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series Molecular Systems Biology
spelling doaj-art-286c8d758b2e4464b99c7cda8cc0b0ab2025-08-20T03:43:34ZengSpringer NatureMolecular Systems Biology1744-42922014-11-01101112110.15252/msb.20145508Rapid neurogenesis through transcriptional activation in human stem cellsVolker Busskamp0Nathan E Lewis1Patrick Guye2Alex HM Ng3Seth L Shipman4Susan M Byrne5Neville E Sanjana6Jernej Murn7Yinqing Li8Shangzhong Li9Michael Stadler10Ron Weiss11George M Church12Department of Genetics, Harvard Medical SchoolDepartment of Genetics, Harvard Medical SchoolDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Genetics, Harvard Medical SchoolDepartment of Genetics, Harvard Medical SchoolDepartment of Genetics, Harvard Medical SchoolBroad Institute of MIT and Harvard, Cambridge CenterDepartment of Cell Biology, Harvard Medical SchoolDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Bioengineering, University of CaliforniaFriedrich Miescher Institute for Biomedical ResearchDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Genetics, Harvard Medical SchoolAbstract Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yields. Here, we overexpressed two Neurogenin transcription factors in human‐induced pluripotent stem cells and obtained neurons with bipolar morphology in 4 days, at greater than 90% purity. The high purity enabled mRNA and microRNA expression profiling during neurogenesis, thus revealing the genetic programs involved in the rapid transition from stem cell to neuron. The resulting cells exhibited transcriptional, morphological and functional signatures of differentiated neurons, with greatest transcriptional similarity to prenatal human brain samples. Our analysis revealed a network of key transcription factors and microRNAs that promoted loss of pluripotency and rapid neurogenesis via progenitor states. Perturbations of key transcription factors affected homogeneity and phenotypic properties of the resulting neurons, suggesting that a systems‐level view of the molecular biology of differentiation may guide subsequent manipulation of human stem cells to rapidly obtain diverse neuronal types.https://doi.org/10.15252/msb.20145508gene regulatory networksmicroRNAsneurogenesisstem cell differentiationtranscriptomics
spellingShingle Volker Busskamp
Nathan E Lewis
Patrick Guye
Alex HM Ng
Seth L Shipman
Susan M Byrne
Neville E Sanjana
Jernej Murn
Yinqing Li
Shangzhong Li
Michael Stadler
Ron Weiss
George M Church
Rapid neurogenesis through transcriptional activation in human stem cells
Molecular Systems Biology
gene regulatory networks
microRNAs
neurogenesis
stem cell differentiation
transcriptomics
title Rapid neurogenesis through transcriptional activation in human stem cells
title_full Rapid neurogenesis through transcriptional activation in human stem cells
title_fullStr Rapid neurogenesis through transcriptional activation in human stem cells
title_full_unstemmed Rapid neurogenesis through transcriptional activation in human stem cells
title_short Rapid neurogenesis through transcriptional activation in human stem cells
title_sort rapid neurogenesis through transcriptional activation in human stem cells
topic gene regulatory networks
microRNAs
neurogenesis
stem cell differentiation
transcriptomics
url https://doi.org/10.15252/msb.20145508
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