绘制人类细胞的遗传全景图。
Mapping the Genetic Landscape of Human Cells.
机构信息
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA; California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, CA 94158, USA.
Department of Bioengineering and ChEM-H, Stanford University, Stanford, CA 94305, USA.
出版信息
Cell. 2018 Aug 9;174(4):953-967.e22. doi: 10.1016/j.cell.2018.06.010. Epub 2018 Jul 19.
Seminal yeast studies have established the value of comprehensively mapping genetic interactions (GIs) for inferring gene function. Efforts in human cells using focused gene sets underscore the utility of this approach, but the feasibility of generating large-scale, diverse human GI maps remains unresolved. We developed a CRISPR interference platform for large-scale quantitative mapping of human GIs. We systematically perturbed 222,784 gene pairs in two cancer cell lines. The resultant maps cluster functionally related genes, assigning function to poorly characterized genes, including TMEM261, a new electron transport chain component. Individual GIs pinpoint unexpected relationships between pathways, exemplified by a specific cholesterol biosynthesis intermediate whose accumulation induces deoxynucleotide depletion, causing replicative DNA damage and a synthetic-lethal interaction with the ATR/9-1-1 DNA repair pathway. Our map provides a broad resource, establishes GI maps as a high-resolution tool for dissecting gene function, and serves as a blueprint for mapping the genetic landscape of human cells.
酵母的研究成果已经证实，全面绘制遗传相互作用（Genetic Interactions，GIs）图谱对于推断基因功能具有重要价值。在人类细胞中使用有针对性的基因集进行的研究强调了这种方法的实用性，但生成大规模、多样化的人类 GI 图谱的可行性仍未得到解决。我们开发了一种用于大规模定量绘制人类 GI 图谱的 CRISPR 干扰平台。我们在两种癌细胞系中系统地扰动了 222,784 对基因对。生成的图谱将功能相关的基因聚类在一起，为功能未知的基因赋予功能，包括 TMEM261，这是一种新的电子传递链组成部分。个别 GI 图谱揭示了不同通路之间意想不到的关系，例如胆固醇生物合成中间产物的积累会导致脱氧核苷酸耗竭，从而导致复制性 DNA 损伤，并与 ATR/9-1-1 DNA 修复通路产生合成致死性相互作用。我们的图谱提供了广泛的资源，将 GI 图谱确立为解析基因功能的高分辨率工具，并为绘制人类细胞的遗传景观蓝图奠定了基础。
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