Loss of tet enzymes compromises proper differentiation of embryonic stem cells

Meelad M. Dawlaty; Achim Breiling; Thuc Le; M. Inmaculada Barrasa; Günter Raddatz; Qing Gao; Benjamin E. Powell; Albert W. Cheng; Kym F. Faull; Frank Lyko; Rudolf Jaenisch

doi:10.1016/j.devcel.2014.03.003

Loss of tet enzymes compromises proper differentiation of embryonic stem cells

Meelad M. Dawlaty, Achim Breiling, Thuc Le, M. Inmaculada Barrasa, Günter Raddatz, Qing Gao, Benjamin E. Powell, Albert W. Cheng, Kym F. Faull, Frank Lyko, Rudolf Jaenisch

Research output: Contribution to journal › Article › peer-review

248 Scopus citations

Abstract

Tet enzymes (Tet1/2/3) convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and aredynamically expressed during development. Whereas loss of individual Tet enzymes or combined deficiency of Tet1/2 allows for embryogenesis, the effect of complete loss of Tet activity and 5hmC marks in development is not established. We have generated Tet1/2/3 triple-knockout (TKO) mouse embryonic stem cells (ESCs) and examined their developmental potential. Combined deficiency of all three Tets depleted 5hmC and impaired ESC differentiation, as seen in poorly differentiated TKO embryoid bodies (EBs) and teratomas. Consistent with impaired differentiation, TKO ESCs contributed poorly to chimeric embryos, a defect rescued by Tet1 reexpression, and could not support embryonic development. Global gene-expression and methylome analyses of TKO EBs revealed promoter hypermethylation and deregulation of genes implicated in embryonic development and differentiation. These findings suggest a requirement for Tet- and 5hmC-mediated DNA demethylation in proper regulation of gene expression during ESC differentiation and development.

Original language	English (US)
Pages (from-to)	102-111
Number of pages	10
Journal	Developmental cell
Volume	29
Issue number	1
DOIs	https://doi.org/10.1016/j.devcel.2014.03.003
State	Published - Apr 14 2014
Externally published	Yes

ASJC Scopus subject areas

Molecular Biology
General Biochemistry, Genetics and Molecular Biology
Developmental Biology
Cell Biology

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10.1016/j.devcel.2014.03.003

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@article{401538fbafac4752b2563409201273ba,

title = "Loss of tet enzymes compromises proper differentiation of embryonic stem cells",

abstract = "Tet enzymes (Tet1/2/3) convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and aredynamically expressed during development. Whereas loss of individual Tet enzymes or combined deficiency of Tet1/2 allows for embryogenesis, the effect of complete loss of Tet activity and 5hmC marks in development is not established. We have generated Tet1/2/3 triple-knockout (TKO) mouse embryonic stem cells (ESCs) and examined their developmental potential. Combined deficiency of all three Tets depleted 5hmC and impaired ESC differentiation, as seen in poorly differentiated TKO embryoid bodies (EBs) and teratomas. Consistent with impaired differentiation, TKO ESCs contributed poorly to chimeric embryos, a defect rescued by Tet1 reexpression, and could not support embryonic development. Global gene-expression and methylome analyses of TKO EBs revealed promoter hypermethylation and deregulation of genes implicated in embryonic development and differentiation. These findings suggest a requirement for Tet- and 5hmC-mediated DNA demethylation in proper regulation of gene expression during ESC differentiation and development.",

author = "Dawlaty, {Meelad M.} and Achim Breiling and Thuc Le and Barrasa, {M. Inmaculada} and G{\"u}nter Raddatz and Qing Gao and Powell, {Benjamin E.} and Cheng, {Albert W.} and Faull, {Kym F.} and Frank Lyko and Rudolf Jaenisch",

note = "Funding Information: We thank Kibibi Ganz, Dongdong Fu, Ruth Flannery, Linyu Shi, Hui Yang, Ping Xu, and Raaji Alagappan for help with animal husbandry, histology, and blastocyst injections. We are also grateful to Tayyeb Adil, John Cassady, Thorold Theunissen, and Jianlong Wang for plasmids, Alex Yoon for help with mass spectrometry, Tanja Musch for 454 sequencing, and the DKFZ Genomics and Proteomics Core Facility for Illumina sequencing services. M.M.D. is a Damon Runyon Postdoctoral Fellow. B.E.P. is supported by a PhD fellowship from the Boehringer Ingelheim Fonds. A.W.C. is supported by a Croucher scholarship. T.L. is supported by a UCLA Molecular, Cellular and Neurobiology training grant, UCLA Mental Retardation training grant, and Eugene V. Cota-Robles fellowship. Work in F.L.{\textquoteright}s lab was supported by a grant from the Deutsche Forschungsgemeinschaft (SPP 1463). R.J. is funded by NIH grants 5-R01-HDO45022 and 5-R37-CA084198 and the Simons Foundation. R.J. is an advisor to Stemgent and cofounder of Fate Therapeutics. ",

year = "2014",

month = apr,

day = "14",

doi = "10.1016/j.devcel.2014.03.003",

language = "English (US)",

volume = "29",

pages = "102--111",

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T1 - Loss of tet enzymes compromises proper differentiation of embryonic stem cells

AU - Dawlaty, Meelad M.

AU - Breiling, Achim

AU - Le, Thuc

AU - Barrasa, M. Inmaculada

AU - Raddatz, Günter

AU - Gao, Qing

AU - Powell, Benjamin E.

AU - Cheng, Albert W.

AU - Faull, Kym F.

AU - Lyko, Frank

AU - Jaenisch, Rudolf

N1 - Funding Information: We thank Kibibi Ganz, Dongdong Fu, Ruth Flannery, Linyu Shi, Hui Yang, Ping Xu, and Raaji Alagappan for help with animal husbandry, histology, and blastocyst injections. We are also grateful to Tayyeb Adil, John Cassady, Thorold Theunissen, and Jianlong Wang for plasmids, Alex Yoon for help with mass spectrometry, Tanja Musch for 454 sequencing, and the DKFZ Genomics and Proteomics Core Facility for Illumina sequencing services. M.M.D. is a Damon Runyon Postdoctoral Fellow. B.E.P. is supported by a PhD fellowship from the Boehringer Ingelheim Fonds. A.W.C. is supported by a Croucher scholarship. T.L. is supported by a UCLA Molecular, Cellular and Neurobiology training grant, UCLA Mental Retardation training grant, and Eugene V. Cota-Robles fellowship. Work in F.L.’s lab was supported by a grant from the Deutsche Forschungsgemeinschaft (SPP 1463). R.J. is funded by NIH grants 5-R01-HDO45022 and 5-R37-CA084198 and the Simons Foundation. R.J. is an advisor to Stemgent and cofounder of Fate Therapeutics.

PY - 2014/4/14

Y1 - 2014/4/14

N2 - Tet enzymes (Tet1/2/3) convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and aredynamically expressed during development. Whereas loss of individual Tet enzymes or combined deficiency of Tet1/2 allows for embryogenesis, the effect of complete loss of Tet activity and 5hmC marks in development is not established. We have generated Tet1/2/3 triple-knockout (TKO) mouse embryonic stem cells (ESCs) and examined their developmental potential. Combined deficiency of all three Tets depleted 5hmC and impaired ESC differentiation, as seen in poorly differentiated TKO embryoid bodies (EBs) and teratomas. Consistent with impaired differentiation, TKO ESCs contributed poorly to chimeric embryos, a defect rescued by Tet1 reexpression, and could not support embryonic development. Global gene-expression and methylome analyses of TKO EBs revealed promoter hypermethylation and deregulation of genes implicated in embryonic development and differentiation. These findings suggest a requirement for Tet- and 5hmC-mediated DNA demethylation in proper regulation of gene expression during ESC differentiation and development.

AB - Tet enzymes (Tet1/2/3) convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and aredynamically expressed during development. Whereas loss of individual Tet enzymes or combined deficiency of Tet1/2 allows for embryogenesis, the effect of complete loss of Tet activity and 5hmC marks in development is not established. We have generated Tet1/2/3 triple-knockout (TKO) mouse embryonic stem cells (ESCs) and examined their developmental potential. Combined deficiency of all three Tets depleted 5hmC and impaired ESC differentiation, as seen in poorly differentiated TKO embryoid bodies (EBs) and teratomas. Consistent with impaired differentiation, TKO ESCs contributed poorly to chimeric embryos, a defect rescued by Tet1 reexpression, and could not support embryonic development. Global gene-expression and methylome analyses of TKO EBs revealed promoter hypermethylation and deregulation of genes implicated in embryonic development and differentiation. These findings suggest a requirement for Tet- and 5hmC-mediated DNA demethylation in proper regulation of gene expression during ESC differentiation and development.

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ER -

Loss of tet enzymes compromises proper differentiation of embryonic stem cells

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