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Characterization of a novel cell penetrating peptide derived from human Oct4
BACKGROUND: Oct4 is a transcription factor that plays a major role for the preservation of the pluripotent state in embryonic stem cells as well as for efficient reprogramming of somatic cells to induced pluripotent stem cells (iPSC) or other progenitors. Protein-based reprogramming methods mainly r...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4230757/ https://www.ncbi.nlm.nih.gov/pubmed/25408881 http://dx.doi.org/10.1186/2045-9769-3-2 |
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author | Harreither, Eva Rydberg, Hanna A Åmand, Helene L Jadhav, Vaibhav Fliedl, Lukas Benda, Christina Esteban, Miguel A Pei, Duanqing Borth, Nicole Grillari-Voglauer, Regina Hommerding, Oliver Edenhofer, Frank Nordén, Bengt Grillari, Johannes |
author_facet | Harreither, Eva Rydberg, Hanna A Åmand, Helene L Jadhav, Vaibhav Fliedl, Lukas Benda, Christina Esteban, Miguel A Pei, Duanqing Borth, Nicole Grillari-Voglauer, Regina Hommerding, Oliver Edenhofer, Frank Nordén, Bengt Grillari, Johannes |
author_sort | Harreither, Eva |
collection | PubMed |
description | BACKGROUND: Oct4 is a transcription factor that plays a major role for the preservation of the pluripotent state in embryonic stem cells as well as for efficient reprogramming of somatic cells to induced pluripotent stem cells (iPSC) or other progenitors. Protein-based reprogramming methods mainly rely on the addition of a fused cell penetrating peptide. This study describes that Oct4 inherently carries a protein transduction domain, which can translocate into human and mouse cells. RESULTS: A 16 amino acid peptide representing the third helix of the human Oct4 homeodomain, referred to as Oct4 protein transduction domain (Oct4-PTD), can internalize in mammalian cells upon conjugation to a fluorescence moiety thereby acting as a cell penetrating peptide (CPP). The cellular distribution of Oct4-PTD shows diffuse cytosolic and nuclear staining, whereas penetratin is strictly localized to a punctuate pattern in the cytoplasm. By using a Cre/loxP-based reporter system, we show that this peptide also drives translocation of a functionally active Oct4-PTD-Cre-fusion protein. We further provide evidence for translocation of full length Oct4 into human and mouse cell lines without the addition of any kind of cationic fusion tag. Finally, physico-chemical properties of the novel CPP are characterized, showing that in contrast to penetratin a helical structure of Oct4-PTD is only observed if the FITC label is present on the N-terminus of the peptide. CONCLUSIONS: Oct4 is a key transcription factor in stem cell research and cellular reprogramming. Since it has been shown that recombinant Oct4 fused to a cationic fusion tag can drive generation of iPSCs, our finding might contribute to further development of protein-based methods to generate iPSCs. Moreover, our data support the idea that transcription factors might be part of an alternative paracrine signalling pathway, where the proteins are transferred to neighbouring cells thereby actively changing the behaviour of the recipient cell. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi: 10.1186/2045-9769-3-2) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4230757 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-42307572014-11-18 Characterization of a novel cell penetrating peptide derived from human Oct4 Harreither, Eva Rydberg, Hanna A Åmand, Helene L Jadhav, Vaibhav Fliedl, Lukas Benda, Christina Esteban, Miguel A Pei, Duanqing Borth, Nicole Grillari-Voglauer, Regina Hommerding, Oliver Edenhofer, Frank Nordén, Bengt Grillari, Johannes Cell Regen Research BACKGROUND: Oct4 is a transcription factor that plays a major role for the preservation of the pluripotent state in embryonic stem cells as well as for efficient reprogramming of somatic cells to induced pluripotent stem cells (iPSC) or other progenitors. Protein-based reprogramming methods mainly rely on the addition of a fused cell penetrating peptide. This study describes that Oct4 inherently carries a protein transduction domain, which can translocate into human and mouse cells. RESULTS: A 16 amino acid peptide representing the third helix of the human Oct4 homeodomain, referred to as Oct4 protein transduction domain (Oct4-PTD), can internalize in mammalian cells upon conjugation to a fluorescence moiety thereby acting as a cell penetrating peptide (CPP). The cellular distribution of Oct4-PTD shows diffuse cytosolic and nuclear staining, whereas penetratin is strictly localized to a punctuate pattern in the cytoplasm. By using a Cre/loxP-based reporter system, we show that this peptide also drives translocation of a functionally active Oct4-PTD-Cre-fusion protein. We further provide evidence for translocation of full length Oct4 into human and mouse cell lines without the addition of any kind of cationic fusion tag. Finally, physico-chemical properties of the novel CPP are characterized, showing that in contrast to penetratin a helical structure of Oct4-PTD is only observed if the FITC label is present on the N-terminus of the peptide. CONCLUSIONS: Oct4 is a key transcription factor in stem cell research and cellular reprogramming. Since it has been shown that recombinant Oct4 fused to a cationic fusion tag can drive generation of iPSCs, our finding might contribute to further development of protein-based methods to generate iPSCs. Moreover, our data support the idea that transcription factors might be part of an alternative paracrine signalling pathway, where the proteins are transferred to neighbouring cells thereby actively changing the behaviour of the recipient cell. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi: 10.1186/2045-9769-3-2) contains supplementary material, which is available to authorized users. BioMed Central 2014-01-31 /pmc/articles/PMC4230757/ /pubmed/25408881 http://dx.doi.org/10.1186/2045-9769-3-2 Text en © Harreither et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Harreither, Eva Rydberg, Hanna A Åmand, Helene L Jadhav, Vaibhav Fliedl, Lukas Benda, Christina Esteban, Miguel A Pei, Duanqing Borth, Nicole Grillari-Voglauer, Regina Hommerding, Oliver Edenhofer, Frank Nordén, Bengt Grillari, Johannes Characterization of a novel cell penetrating peptide derived from human Oct4 |
title | Characterization of a novel cell penetrating peptide derived from human Oct4 |
title_full | Characterization of a novel cell penetrating peptide derived from human Oct4 |
title_fullStr | Characterization of a novel cell penetrating peptide derived from human Oct4 |
title_full_unstemmed | Characterization of a novel cell penetrating peptide derived from human Oct4 |
title_short | Characterization of a novel cell penetrating peptide derived from human Oct4 |
title_sort | characterization of a novel cell penetrating peptide derived from human oct4 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4230757/ https://www.ncbi.nlm.nih.gov/pubmed/25408881 http://dx.doi.org/10.1186/2045-9769-3-2 |
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