Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects
Defining the genes that are essential for cellular proliferation is critical for understanding organismal development and identifying high-value targets for disease therapies. However, the requirements for cell-cycle progression in human cells remain incompletely understood. To elucidate the consequ...
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| Format: | Article |
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Elsevier BV
2018
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| Online Access: | http://hdl.handle.net/1721.1/116347 https://orcid.org/0000-0001-6283-9168 https://orcid.org/0000-0002-3829-5612 |
| _version_ | 1826216080054419456 |
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| author | McKinley, Kara Lavidge Cheeseman, Iain M |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology McKinley, Kara Lavidge Cheeseman, Iain M |
| author_sort | McKinley, Kara Lavidge |
| collection | MIT |
| description | Defining the genes that are essential for cellular proliferation is critical for understanding organismal development and identifying high-value targets for disease therapies. However, the requirements for cell-cycle progression in human cells remain incompletely understood. To elucidate the consequences of acute and chronic elimination of cell-cycle proteins, we generated and characterized inducible CRISPR/Cas9 knockout human cell lines targeting 209 genes involved in diverse cell-cycle processes. We performed single-cell microscopic analyses to systematically establish the effects of the knockouts on subcellular architecture. To define variations in cell-cycle requirements between cultured cell lines, we generated knockouts across cell lines of diverse origins. We demonstrate that p53 modulates the phenotype of specific cell-cycle defects through distinct mechanisms, depending on the defect. This work provides a resource to broadly facilitate robust and long-term depletion of cell-cycle proteins and reveals insights into the requirements for cell-cycle progression. Keywords:
kinetochore; centromere; mitosis; DNA replication; spindle; microtubule; multipolarity; p53; CRISPR/Cas9 |
| first_indexed | 2024-09-23T16:42:00Z |
| format | Article |
| id | mit-1721.1/116347 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T16:42:00Z |
| publishDate | 2018 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | mit-1721.1/1163472022-10-03T07:41:03Z Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects McKinley, Kara Lavidge Cheeseman, Iain M Massachusetts Institute of Technology. Department of Biology McKinley, Kara Lavidge Cheeseman, Iain M Defining the genes that are essential for cellular proliferation is critical for understanding organismal development and identifying high-value targets for disease therapies. However, the requirements for cell-cycle progression in human cells remain incompletely understood. To elucidate the consequences of acute and chronic elimination of cell-cycle proteins, we generated and characterized inducible CRISPR/Cas9 knockout human cell lines targeting 209 genes involved in diverse cell-cycle processes. We performed single-cell microscopic analyses to systematically establish the effects of the knockouts on subcellular architecture. To define variations in cell-cycle requirements between cultured cell lines, we generated knockouts across cell lines of diverse origins. We demonstrate that p53 modulates the phenotype of specific cell-cycle defects through distinct mechanisms, depending on the defect. This work provides a resource to broadly facilitate robust and long-term depletion of cell-cycle proteins and reveals insights into the requirements for cell-cycle progression. Keywords: kinetochore; centromere; mitosis; DNA replication; spindle; microtubule; multipolarity; p53; CRISPR/Cas9 2018-06-15T18:09:28Z 2018-06-15T18:09:28Z 2017-02 2016-10 2018-06-14T13:45:06Z Article http://purl.org/eprint/type/JournalArticle 1534-5807 1878-1551 http://hdl.handle.net/1721.1/116347 McKinley, Kara L. and Iain M. Cheeseman. “Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of P53-Dependent Responses to Cell-Cycle Defects.” Developmental Cell 40, 4 (February 2017): 405–420 © 2017 Elsevier Inc https://orcid.org/0000-0001-6283-9168 https://orcid.org/0000-0002-3829-5612 http://dx.doi.org/10.1016/J.DEVCEL.2017.01.012 Developmental Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier BV PMC |
| spellingShingle | McKinley, Kara Lavidge Cheeseman, Iain M Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects |
| title | Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects |
| title_full | Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects |
| title_fullStr | Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects |
| title_full_unstemmed | Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects |
| title_short | Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects |
| title_sort | large scale analysis of crispr cas9 cell cycle knockouts reveals the diversity of p53 dependent responses to cell cycle defects |
| url | http://hdl.handle.net/1721.1/116347 https://orcid.org/0000-0001-6283-9168 https://orcid.org/0000-0002-3829-5612 |
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