Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects.
Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects.
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DOI：
10.1016/j.devcel.2017.01.012
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发表时间：
2017-02-27
影响因子：
11.8
通讯作者：
Cheeseman IM
中科院分区：
文献类型：
--
作者：
McKinley KL;Cheeseman IM
定义细胞增殖所必需的基因对于理解有机体发育和确定疾病治疗的高价值靶点至关重要。然而，人类细胞细胞周期进展的要求仍未完全了解。为了阐明细胞周期蛋白急性和慢性消除的后果，我们生成并表征了针对参与不同细胞周期过程的 209 个基因的可诱导 CRISPR/Cas9 敲除人类细胞系。我们进行了单细胞显微镜分析，以系统地确定敲除对亚细胞结构的影响。为了定义培养细胞系之间细胞周期要求的差异，我们对不同来源的细胞系进行了敲除。我们证明 p53 根据缺陷通过不同的机制调节特定细胞周期缺陷的表型。这项工作提供了广泛促进细胞周期蛋白的稳健和长期消耗的资源，并揭示了对细胞周期进展的要求的见解。 McKinley 和 Cheeseman 报告了大约 500 个可诱导 CRISPR/Cas9 敲除细胞系的资源，这些细胞系针对参与细胞周期过程的 209 个基因。定义与这些细胞系相关的相应表型并对不同细胞类型进行比较揭示了 p53 通路对特定细胞周期缺陷的不同反应。
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. McKinley and Cheeseman report a resource of ~500 inducible CRISPR/Cas9 knockout cell lines targeting 209 genes involved in cell-cycle processes. Defining the corresponding phenotypes associated with these cell lines and comparing them across diverse cell types reveals differential responses of the p53 pathway to specific cell-cycle defects.