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Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency

Revealing the molecular events associated with reprogramming different somatic cell types to pluripotency is critical for understanding the characteristics of induced pluripotent stem cell (iPSC) therapeutic derivatives. Inducible reprogramming factor transgenic cells or animals—designated as second...

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Autores principales: Elbaz, Judith, Puri, Mira C., Faiz, Maryam, Bang, K. W. Annie, Nguyen, Lena, Makovoz, Bar, Gertsenstein, Marina, Hussein, Samer M. I., Zandstra, Peter W., Briollais, Laurent, Shakiba, Nika, Nagy, Andras
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9894234/
https://www.ncbi.nlm.nih.gov/pubmed/36454756
http://dx.doi.org/10.1073/pnas.2207824119
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author Elbaz, Judith
Puri, Mira C.
Faiz, Maryam
Bang, K. W. Annie
Nguyen, Lena
Makovoz, Bar
Gertsenstein, Marina
Hussein, Samer M. I.
Zandstra, Peter W.
Briollais, Laurent
Shakiba, Nika
Nagy, Andras
author_facet Elbaz, Judith
Puri, Mira C.
Faiz, Maryam
Bang, K. W. Annie
Nguyen, Lena
Makovoz, Bar
Gertsenstein, Marina
Hussein, Samer M. I.
Zandstra, Peter W.
Briollais, Laurent
Shakiba, Nika
Nagy, Andras
author_sort Elbaz, Judith
collection PubMed
description Revealing the molecular events associated with reprogramming different somatic cell types to pluripotency is critical for understanding the characteristics of induced pluripotent stem cell (iPSC) therapeutic derivatives. Inducible reprogramming factor transgenic cells or animals—designated as secondary (2°) reprogramming systems—not only provide excellent experimental tools for such studies but also offer a strategy to study the variances in cellular reprogramming outcomes due to different in vitro and in vivo environments. To make such studies less cumbersome, it is desirable to have a variety of efficient reprogrammable mouse systems to induce successful mass reprogramming in somatic cell types. Here, we report the development of two transgenic mouse lines from which 2° cells reprogram with unprecedented efficiency. These systems were derived by exposing primary reprogramming cells containing doxycycline-inducible Yamanaka factor expression to a transient interruption in transgene expression, resulting in selection for a subset of clones with robust transgene response. These systems also include reporter genes enabling easy readout of endogenous Oct4 activation (GFP), indicative of pluripotency, and reprogramming transgene expression (mCherry). Notably, somatic cells derived from various fetal and adult tissues from these 2° mouse lines gave rise to highly efficient and rapid reprogramming, with transgene-independent iPSC colonies emerging as early as 1 wk after induction. These mouse lines serve as a powerful tool to explore sources of variability in reprogramming and the mechanistic underpinnings of efficient reprogramming systems.
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spelling pubmed-98942342023-02-03 Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency Elbaz, Judith Puri, Mira C. Faiz, Maryam Bang, K. W. Annie Nguyen, Lena Makovoz, Bar Gertsenstein, Marina Hussein, Samer M. I. Zandstra, Peter W. Briollais, Laurent Shakiba, Nika Nagy, Andras Proc Natl Acad Sci U S A Biological Sciences Revealing the molecular events associated with reprogramming different somatic cell types to pluripotency is critical for understanding the characteristics of induced pluripotent stem cell (iPSC) therapeutic derivatives. Inducible reprogramming factor transgenic cells or animals—designated as secondary (2°) reprogramming systems—not only provide excellent experimental tools for such studies but also offer a strategy to study the variances in cellular reprogramming outcomes due to different in vitro and in vivo environments. To make such studies less cumbersome, it is desirable to have a variety of efficient reprogrammable mouse systems to induce successful mass reprogramming in somatic cell types. Here, we report the development of two transgenic mouse lines from which 2° cells reprogram with unprecedented efficiency. These systems were derived by exposing primary reprogramming cells containing doxycycline-inducible Yamanaka factor expression to a transient interruption in transgene expression, resulting in selection for a subset of clones with robust transgene response. These systems also include reporter genes enabling easy readout of endogenous Oct4 activation (GFP), indicative of pluripotency, and reprogramming transgene expression (mCherry). Notably, somatic cells derived from various fetal and adult tissues from these 2° mouse lines gave rise to highly efficient and rapid reprogramming, with transgene-independent iPSC colonies emerging as early as 1 wk after induction. These mouse lines serve as a powerful tool to explore sources of variability in reprogramming and the mechanistic underpinnings of efficient reprogramming systems. National Academy of Sciences 2022-12-01 2022-12-06 /pmc/articles/PMC9894234/ /pubmed/36454756 http://dx.doi.org/10.1073/pnas.2207824119 Text en Copyright © 2022 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Elbaz, Judith
Puri, Mira C.
Faiz, Maryam
Bang, K. W. Annie
Nguyen, Lena
Makovoz, Bar
Gertsenstein, Marina
Hussein, Samer M. I.
Zandstra, Peter W.
Briollais, Laurent
Shakiba, Nika
Nagy, Andras
Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency
title Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency
title_full Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency
title_fullStr Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency
title_full_unstemmed Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency
title_short Highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency
title_sort highly efficient reprogrammable mouse lines with integrated reporters to track the route to pluripotency
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9894234/
https://www.ncbi.nlm.nih.gov/pubmed/36454756
http://dx.doi.org/10.1073/pnas.2207824119
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