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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2018-10-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/36045 |
| _version_ | 1811253283504783360 |
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| author | Ashley RG Libby David A Joy Po-Lin So Mohammad A Mandegar Jonathon M Muncie Federico N Mendoza-Camacho Valerie M Weaver Bruce R Conklin Todd C McDevitt |
| author_facet | Ashley RG Libby David A Joy Po-Lin So Mohammad A Mandegar Jonathon M Muncie Federico N Mendoza-Camacho Valerie M Weaver Bruce R Conklin Todd C McDevitt |
| author_sort | Ashley RG Libby |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-12T16:47:35Z |
| format | Article |
| id | doaj.art-f96a0f62c79f4d14993cf3909712610d |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:47:35Z |
| publishDate | 2018-10-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-f96a0f62c79f4d14993cf3909712610d2022-12-22T03:24:31ZengeLife Sciences Publications LtdeLife2050-084X2018-10-01710.7554/eLife.36045Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interferenceAshley RG Libby0https://orcid.org/0000-0002-8139-8844David A Joy1Po-Lin So2Mohammad A Mandegar3Jonathon M Muncie4Federico N Mendoza-Camacho5Valerie M Weaver6Bruce R Conklin7Todd C McDevitt8https://orcid.org/0000-0002-8905-4931Gladstone Institute of Cardiovascular Disease, San Francisco, United States; Developmental and Stem Cell Biology Graduate Program, University of California San Francisco, San Francisco, United StatesGladstone Institute of Cardiovascular Disease, San Francisco, United States; Graduate Program in Bioengineering, University of California Berkeley, University of California San Francisco, San Francisco, United StatesGladstone Institute of Cardiovascular Disease, San Francisco, United StatesGladstone Institute of Cardiovascular Disease, San Francisco, United StatesGraduate Program in Bioengineering, University of California Berkeley, University of California San Francisco, San Francisco, United States; Department of Surgery, Center for Bioengineering and Tissue Regeneration, University of California, San Francisco, United StatesGladstone Institute of Cardiovascular Disease, San Francisco, United StatesDepartment of Surgery, Center for Bioengineering and Tissue Regeneration, University of California, San Francisco, United StatesGladstone Institute of Cardiovascular Disease, San Francisco, United States; Department of Medicine, Division of Genomic Medicine, University of California, San Francisco, United StatesGladstone Institute of Cardiovascular Disease, San Francisco, United States; Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, United StatesMorphogenesis 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.https://elifesciences.org/articles/36045pluripotent stem cellsmorphogenesisbio-engineering |
| spellingShingle | Ashley RG Libby David A Joy Po-Lin So Mohammad A Mandegar Jonathon M Muncie Federico N Mendoza-Camacho Valerie M Weaver Bruce R Conklin Todd C McDevitt Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference eLife pluripotent stem cells morphogenesis bio-engineering |
| 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 | pluripotent stem cells morphogenesis bio-engineering |
| url | https://elifesciences.org/articles/36045 |
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