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Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference

Morphogenesis involves interactions of asymmetric cell populations to form complex multicellular patterns and structures comprised of distinct cell types. However, current methods to model morphogenic events lack control over cell-type co-emergence and offer little capability to selectively perturb...

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Autores principales: Libby, Ashley RG, Joy, David A, So, Po-Lin, Mandegar, Mohammad A, Muncie, Jonathon M, Mendoza-Camacho, Federico N, Weaver, Valerie M, Conklin, Bruce R, McDevitt, Todd C
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6177255/
https://www.ncbi.nlm.nih.gov/pubmed/30298816
http://dx.doi.org/10.7554/eLife.36045
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author Libby, Ashley RG
Joy, David A
So, Po-Lin
Mandegar, Mohammad A
Muncie, Jonathon M
Mendoza-Camacho, Federico N
Weaver, Valerie M
Conklin, Bruce R
McDevitt, Todd C
author_facet Libby, Ashley RG
Joy, David A
So, Po-Lin
Mandegar, Mohammad A
Muncie, Jonathon M
Mendoza-Camacho, Federico N
Weaver, Valerie M
Conklin, Bruce R
McDevitt, Todd C
author_sort Libby, Ashley RG
collection PubMed
description Morphogenesis involves interactions of asymmetric cell populations to form complex multicellular patterns and structures comprised of distinct cell types. However, current methods to model morphogenic events lack control over cell-type co-emergence and offer little capability to selectively perturb specific cell subpopulations. Our in vitro system interrogates cell-cell interactions and multicellular organization within human induced pluripotent stem cell (hiPSC) colonies. We examined effects of induced mosaic knockdown of molecular regulators of cortical tension (ROCK1) and cell-cell adhesion (CDH1) with CRISPR interference. Mosaic knockdown of ROCK1 or CDH1 resulted in differential patterning within hiPSC colonies due to cellular self-organization, while retaining an epithelial pluripotent phenotype. Knockdown induction stimulates a transient wave of differential gene expression within the mixed populations that stabilized in coordination with observed self-organization. Mosaic patterning enables genetic interrogation of emergent multicellular properties, which can facilitate better understanding of the molecular pathways that regulate symmetry-breaking during morphogenesis.
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spelling pubmed-61772552018-10-15 Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference Libby, Ashley RG Joy, David A So, Po-Lin Mandegar, Mohammad A Muncie, Jonathon M Mendoza-Camacho, Federico N Weaver, Valerie M Conklin, Bruce R McDevitt, Todd C eLife Developmental Biology Morphogenesis involves interactions of asymmetric cell populations to form complex multicellular patterns and structures comprised of distinct cell types. However, current methods to model morphogenic events lack control over cell-type co-emergence and offer little capability to selectively perturb specific cell subpopulations. Our in vitro system interrogates cell-cell interactions and multicellular organization within human induced pluripotent stem cell (hiPSC) colonies. We examined effects of induced mosaic knockdown of molecular regulators of cortical tension (ROCK1) and cell-cell adhesion (CDH1) with CRISPR interference. Mosaic knockdown of ROCK1 or CDH1 resulted in differential patterning within hiPSC colonies due to cellular self-organization, while retaining an epithelial pluripotent phenotype. Knockdown induction stimulates a transient wave of differential gene expression within the mixed populations that stabilized in coordination with observed self-organization. Mosaic patterning enables genetic interrogation of emergent multicellular properties, which can facilitate better understanding of the molecular pathways that regulate symmetry-breaking during morphogenesis. eLife Sciences Publications, Ltd 2018-10-09 /pmc/articles/PMC6177255/ /pubmed/30298816 http://dx.doi.org/10.7554/eLife.36045 Text en © 2018, Libby et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Developmental Biology
Libby, Ashley RG
Joy, David A
So, Po-Lin
Mandegar, Mohammad A
Muncie, Jonathon M
Mendoza-Camacho, Federico N
Weaver, Valerie M
Conklin, Bruce R
McDevitt, Todd C
Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference
title Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference
title_full Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference
title_fullStr Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference
title_full_unstemmed Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference
title_short Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference
title_sort spatiotemporal mosaic self-patterning of pluripotent stem cells using crispr interference
topic Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6177255/
https://www.ncbi.nlm.nih.gov/pubmed/30298816
http://dx.doi.org/10.7554/eLife.36045
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