title: Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects.
abstract: 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.
authors.affiliation: Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, MA 02142, USA; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA. Electronic address: kara.l.mckinley@gmail.com.
authors.forename: Kara L
authors.initials: KL
authors.lastname: McKinley
authors.affiliation: Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, MA 02142, USA; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA. Electronic address: icheese@wi.mit.edu.
authors.forename: Iain M
authors.initials: IM
authors.lastname: Cheeseman
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D002453:Cell Cycle / Q000235:genetics*; D016764:Cell Polarity; D049109:Cell Proliferation; D002470:Cell Survival; D016922:Cellular Senescence; D002502:Centrioles / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D055786:Gene Knockout Techniques; D006367:HeLa Cells; D006801:Humans; D010641:Phenotype; D015398:Signal Transduction; D008941:Spindle Apparatus / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D050759:Cyclin-Dependent Kinase Inhibitor p21; D016159:Tumor Suppressor Protein p53
keywords: CRISPR/Cas9; DNA replication; centromere; kinetochore; microtubule; mitosis; multipolarity; p53; spindle