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Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells

Delivery of the transcription factors Oct4, Klf4, Sox2 and c-Myc via integrating viral vectors has been widely employed to generate induced pluripotent stem cell (iPSC) lines from both normal and disease-specific somatic tissues, providing an invaluable resource for medical research and drug develop...

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Autores principales: Sommer, Cesar A., Christodoulou, Constantina, Gianotti-Sommer, Andreia, Shen, Steven S., Sailaja, Badi Sri, Hezroni, Hadas, Spira, Avrum, Meshorer, Eran, Kotton, Darrell N., Mostoslavsky, Gustavo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3522693/
https://www.ncbi.nlm.nih.gov/pubmed/23272148
http://dx.doi.org/10.1371/journal.pone.0051711
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author Sommer, Cesar A.
Christodoulou, Constantina
Gianotti-Sommer, Andreia
Shen, Steven S.
Sailaja, Badi Sri
Hezroni, Hadas
Spira, Avrum
Meshorer, Eran
Kotton, Darrell N.
Mostoslavsky, Gustavo
author_facet Sommer, Cesar A.
Christodoulou, Constantina
Gianotti-Sommer, Andreia
Shen, Steven S.
Sailaja, Badi Sri
Hezroni, Hadas
Spira, Avrum
Meshorer, Eran
Kotton, Darrell N.
Mostoslavsky, Gustavo
author_sort Sommer, Cesar A.
collection PubMed
description Delivery of the transcription factors Oct4, Klf4, Sox2 and c-Myc via integrating viral vectors has been widely employed to generate induced pluripotent stem cell (iPSC) lines from both normal and disease-specific somatic tissues, providing an invaluable resource for medical research and drug development. Residual reprogramming transgene expression from integrated viruses nevertheless alters the biological properties of iPSCs and has been associated with a reduced developmental competence both in vivo and in vitro. We performed transcriptional profiling of mouse iPSC lines before and after excision of a polycistronic lentiviral reprogramming vector to systematically define the overall impact of persistent transgene expression on the molecular features of iPSCs. We demonstrate that residual expression of the Yamanaka factors prevents iPSCs from acquiring the transcriptional program exhibited by embryonic stem cells (ESCs) and that the expression profiles of iPSCs generated with and without c-Myc are indistinguishable. After vector excision, we find 36% of iPSC clones show normal methylation of the Gtl2 region, an imprinted locus that marks ESC-equivalent iPSC lines. Furthermore, we show that the reprogramming factor Klf4 binds to the promoter region of Gtl2. Regardless of Gtl2 methylation status, we find similar endodermal and hepatocyte differentiation potential comparing syngeneic Gtl2(ON) vs Gtl2(OFF) iPSC clones. Our findings provide new insights into the reprogramming process and emphasize the importance of generating iPSCs free of any residual transgene expression.
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spelling pubmed-35226932012-12-27 Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells Sommer, Cesar A. Christodoulou, Constantina Gianotti-Sommer, Andreia Shen, Steven S. Sailaja, Badi Sri Hezroni, Hadas Spira, Avrum Meshorer, Eran Kotton, Darrell N. Mostoslavsky, Gustavo PLoS One Research Article Delivery of the transcription factors Oct4, Klf4, Sox2 and c-Myc via integrating viral vectors has been widely employed to generate induced pluripotent stem cell (iPSC) lines from both normal and disease-specific somatic tissues, providing an invaluable resource for medical research and drug development. Residual reprogramming transgene expression from integrated viruses nevertheless alters the biological properties of iPSCs and has been associated with a reduced developmental competence both in vivo and in vitro. We performed transcriptional profiling of mouse iPSC lines before and after excision of a polycistronic lentiviral reprogramming vector to systematically define the overall impact of persistent transgene expression on the molecular features of iPSCs. We demonstrate that residual expression of the Yamanaka factors prevents iPSCs from acquiring the transcriptional program exhibited by embryonic stem cells (ESCs) and that the expression profiles of iPSCs generated with and without c-Myc are indistinguishable. After vector excision, we find 36% of iPSC clones show normal methylation of the Gtl2 region, an imprinted locus that marks ESC-equivalent iPSC lines. Furthermore, we show that the reprogramming factor Klf4 binds to the promoter region of Gtl2. Regardless of Gtl2 methylation status, we find similar endodermal and hepatocyte differentiation potential comparing syngeneic Gtl2(ON) vs Gtl2(OFF) iPSC clones. Our findings provide new insights into the reprogramming process and emphasize the importance of generating iPSCs free of any residual transgene expression. Public Library of Science 2012-12-14 /pmc/articles/PMC3522693/ /pubmed/23272148 http://dx.doi.org/10.1371/journal.pone.0051711 Text en © 2012 Sommer et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Sommer, Cesar A.
Christodoulou, Constantina
Gianotti-Sommer, Andreia
Shen, Steven S.
Sailaja, Badi Sri
Hezroni, Hadas
Spira, Avrum
Meshorer, Eran
Kotton, Darrell N.
Mostoslavsky, Gustavo
Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells
title Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells
title_full Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells
title_fullStr Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells
title_full_unstemmed Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells
title_short Residual Expression of Reprogramming Factors Affects the Transcriptional Program and Epigenetic Signatures of Induced Pluripotent Stem Cells
title_sort residual expression of reprogramming factors affects the transcriptional program and epigenetic signatures of induced pluripotent stem cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3522693/
https://www.ncbi.nlm.nih.gov/pubmed/23272148
http://dx.doi.org/10.1371/journal.pone.0051711
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