Developmental Cell
Volume 40, Issue 4, 27 February 2017, Pages 405-420.e2
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Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects

https://doi.org/10.1016/j.devcel.2017.01.012Get rights and content
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Highlights

  • Collection of inducible CRISPR knockout cell lines for analysis of cell-cycle genes

  • Systematic analysis of the phenotypes associated with each knockout cell line

  • Inducible knockout cell lines reveal diverse origins of spindle multipolarity

  • The p53 pathway responds differentially to distinct cell-cycle defects

Summary

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

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Present address: University of California, San Francisco, 600 16th Street, San Francisco, CA 94158, USA

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