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Reprogramming Roadblocks Are System Dependent

Since the first generation of induced pluripotent stem cells (iPSCs), several reprogramming systems have been used to study its molecular mechanisms. However, the system of choice largely affects the reprogramming efficiency, influencing our view on the mechanisms. Here, we demonstrate that reprogra...

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Autores principales: Chantzoura, Eleni, Skylaki, Stavroula, Menendez, Sergio, Kim, Shin-Il, Johnsson, Anna, Linnarsson, Sten, Woltjen, Knut, Chambers, Ian, Kaji, Keisuke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4618455/
https://www.ncbi.nlm.nih.gov/pubmed/26278041
http://dx.doi.org/10.1016/j.stemcr.2015.07.007
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author Chantzoura, Eleni
Skylaki, Stavroula
Menendez, Sergio
Kim, Shin-Il
Johnsson, Anna
Linnarsson, Sten
Woltjen, Knut
Chambers, Ian
Kaji, Keisuke
author_facet Chantzoura, Eleni
Skylaki, Stavroula
Menendez, Sergio
Kim, Shin-Il
Johnsson, Anna
Linnarsson, Sten
Woltjen, Knut
Chambers, Ian
Kaji, Keisuke
author_sort Chantzoura, Eleni
collection PubMed
description Since the first generation of induced pluripotent stem cells (iPSCs), several reprogramming systems have been used to study its molecular mechanisms. However, the system of choice largely affects the reprogramming efficiency, influencing our view on the mechanisms. Here, we demonstrate that reprogramming triggered by less efficient polycistronic reprogramming cassettes not only highlights mesenchymal-to-epithelial transition (MET) as a roadblock but also faces more severe difficulties to attain a pluripotent state even post-MET. In contrast, more efficient cassettes can reprogram both wild-type and Nanog(−/−) fibroblasts with comparable efficiencies, routes, and kinetics, unlike the less efficient reprogramming systems. Moreover, we attribute a previously reported variation in the N terminus of KLF4 as a dominant factor underlying these critical differences. Our data establish that some reprogramming roadblocks are system dependent, highlighting the need to pursue mechanistic studies with close attention to the systems to better understand reprogramming.
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spelling pubmed-46184552015-11-24 Reprogramming Roadblocks Are System Dependent Chantzoura, Eleni Skylaki, Stavroula Menendez, Sergio Kim, Shin-Il Johnsson, Anna Linnarsson, Sten Woltjen, Knut Chambers, Ian Kaji, Keisuke Stem Cell Reports Article Since the first generation of induced pluripotent stem cells (iPSCs), several reprogramming systems have been used to study its molecular mechanisms. However, the system of choice largely affects the reprogramming efficiency, influencing our view on the mechanisms. Here, we demonstrate that reprogramming triggered by less efficient polycistronic reprogramming cassettes not only highlights mesenchymal-to-epithelial transition (MET) as a roadblock but also faces more severe difficulties to attain a pluripotent state even post-MET. In contrast, more efficient cassettes can reprogram both wild-type and Nanog(−/−) fibroblasts with comparable efficiencies, routes, and kinetics, unlike the less efficient reprogramming systems. Moreover, we attribute a previously reported variation in the N terminus of KLF4 as a dominant factor underlying these critical differences. Our data establish that some reprogramming roadblocks are system dependent, highlighting the need to pursue mechanistic studies with close attention to the systems to better understand reprogramming. Elsevier 2015-08-13 /pmc/articles/PMC4618455/ /pubmed/26278041 http://dx.doi.org/10.1016/j.stemcr.2015.07.007 Text en © 2015 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Chantzoura, Eleni
Skylaki, Stavroula
Menendez, Sergio
Kim, Shin-Il
Johnsson, Anna
Linnarsson, Sten
Woltjen, Knut
Chambers, Ian
Kaji, Keisuke
Reprogramming Roadblocks Are System Dependent
title Reprogramming Roadblocks Are System Dependent
title_full Reprogramming Roadblocks Are System Dependent
title_fullStr Reprogramming Roadblocks Are System Dependent
title_full_unstemmed Reprogramming Roadblocks Are System Dependent
title_short Reprogramming Roadblocks Are System Dependent
title_sort reprogramming roadblocks are system dependent
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4618455/
https://www.ncbi.nlm.nih.gov/pubmed/26278041
http://dx.doi.org/10.1016/j.stemcr.2015.07.007
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