Multi-omic Analyses Reveal Minimal Impact of the CRISPR-Cas9 Nuclease on Cultured Human Cells.,Journal of Proteome Research

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Multi-omic Analyses Reveal Minimal Impact of the CRISPR-Cas9 Nuclease on Cultured Human Cells.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-02-04 , DOI: 10.1021/acs.jproteome.8b00751
Jiali Qiang _{1,

2} , Zaijun Ma _{1,

2} , Xingxing Xie _{1,

2} , Linyu Shi _{1,

3} , Yang Geng ₁ , Junhao Hu ₁ , Rui Liu ₄ , Nan Liu ₁ , Yaoyang Zhang ₁

Affiliation

The CRISPR-Cas9 system is a genomic editing tool widely used in basic research and under investigation for potential applications in gene therapies for human diseases. To accomplish genomic editing, the system requires the expression of a prokaryotic DNA endonuclease enzyme, Cas9, in host cells. Previous studies have mainly focused on the specificity of Cas9 on the host genome, and thus it is unclear whether this bacterium-derived enzyme affects the protein homeostasis of host cells. Here we applied multi-omic analyses, including transcriptome, proteome, phosphoproteome, Cas9-associated protein interactome, protein synthesis, and histone epigenetic modification, to investigate the cellular response of human cells upon the expression of Cas9. We demonstrate that Cas9 has minimal impact on host cells. Our assessment of intracellular effects of Cas9 paves a path for its broad applications in biological studies and potential clinical translations.

中文翻译：

多组学分析揭示了CRISPR-Cas9核酸酶对培养的人细胞的最小影响。

CRISPR-Cas9系统是一种基因组编辑工具，广泛用于基础研究和正在研究中的人类疾病基因疗法中的潜在应用。为了完成基因组编辑，该系统需要在宿主细胞中表达原核DNA核酸内切酶Cas9。先前的研究主要集中在宿主基因组上Cas9的特异性，因此尚不清楚这种细菌衍生的酶是否影响宿主细胞的蛋白质稳态。在这里，我们应用了多组学分析，包括转录组，蛋白质组，磷酸化蛋白质组，Cas9相关的蛋白质相互作用组，蛋白质合成和组蛋白表观遗传修饰，来研究人类细胞对Cas9表达的细胞反应。我们证明，Cas9对宿主细胞的影响最小。

更新日期：2019-02-07

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