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Arrayed functional genetic screenings in pluripotency reprogramming and differentiation

Thoroughly understanding the molecular mechanisms responsible for the biological properties of pluripotent stem cells, as well as for the processes involved in reprograming, differentiation, and transition between Naïve and Primed pluripotent states, is of great interest in basic and applied researc...

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Autores principales: Panepucci, Rodrigo Alexandre, de Souza Lima, Ildercílio Mota
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6330485/
https://www.ncbi.nlm.nih.gov/pubmed/30635073
http://dx.doi.org/10.1186/s13287-018-1124-6
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author Panepucci, Rodrigo Alexandre
de Souza Lima, Ildercílio Mota
author_facet Panepucci, Rodrigo Alexandre
de Souza Lima, Ildercílio Mota
author_sort Panepucci, Rodrigo Alexandre
collection PubMed
description Thoroughly understanding the molecular mechanisms responsible for the biological properties of pluripotent stem cells, as well as for the processes involved in reprograming, differentiation, and transition between Naïve and Primed pluripotent states, is of great interest in basic and applied research. Although pluripotent cells have been extensively characterized in terms of their transcriptome and miRNome, a comprehensive understanding of how these gene products specifically impact their biology, depends on gain- or loss-of-function experimental approaches capable to systematically interrogate their function. We review all studies carried up to date that used arrayed screening approaches to explore the function of these genetic elements on those biological contexts, using focused or genome-wide genetic libraries. We further discuss the limitations and advantages of approaches based on assays with population-level primary readouts, derived from single-parameter plate readers, or cell-level primary readouts, obtained using multiparametric flow cytometry or quantitative fluorescence microscopy (i.e., high-content screening). Finally, we discuss technical limitation and future perspectives, highlighting how the integration of screening data may lead to major advances in the field of stem cell research and therapy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-1124-6) contains supplementary material, which is available to authorized users.
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spelling pubmed-63304852019-01-16 Arrayed functional genetic screenings in pluripotency reprogramming and differentiation Panepucci, Rodrigo Alexandre de Souza Lima, Ildercílio Mota Stem Cell Res Ther Review Thoroughly understanding the molecular mechanisms responsible for the biological properties of pluripotent stem cells, as well as for the processes involved in reprograming, differentiation, and transition between Naïve and Primed pluripotent states, is of great interest in basic and applied research. Although pluripotent cells have been extensively characterized in terms of their transcriptome and miRNome, a comprehensive understanding of how these gene products specifically impact their biology, depends on gain- or loss-of-function experimental approaches capable to systematically interrogate their function. We review all studies carried up to date that used arrayed screening approaches to explore the function of these genetic elements on those biological contexts, using focused or genome-wide genetic libraries. We further discuss the limitations and advantages of approaches based on assays with population-level primary readouts, derived from single-parameter plate readers, or cell-level primary readouts, obtained using multiparametric flow cytometry or quantitative fluorescence microscopy (i.e., high-content screening). Finally, we discuss technical limitation and future perspectives, highlighting how the integration of screening data may lead to major advances in the field of stem cell research and therapy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-1124-6) contains supplementary material, which is available to authorized users. BioMed Central 2019-01-11 /pmc/articles/PMC6330485/ /pubmed/30635073 http://dx.doi.org/10.1186/s13287-018-1124-6 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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 Review
Panepucci, Rodrigo Alexandre
de Souza Lima, Ildercílio Mota
Arrayed functional genetic screenings in pluripotency reprogramming and differentiation
title Arrayed functional genetic screenings in pluripotency reprogramming and differentiation
title_full Arrayed functional genetic screenings in pluripotency reprogramming and differentiation
title_fullStr Arrayed functional genetic screenings in pluripotency reprogramming and differentiation
title_full_unstemmed Arrayed functional genetic screenings in pluripotency reprogramming and differentiation
title_short Arrayed functional genetic screenings in pluripotency reprogramming and differentiation
title_sort arrayed functional genetic screenings in pluripotency reprogramming and differentiation
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6330485/
https://www.ncbi.nlm.nih.gov/pubmed/30635073
http://dx.doi.org/10.1186/s13287-018-1124-6
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