Haploid Genetic Screens in Human Cells Identify Host Factors Used by Pathogens
Haploid Genetic Screens in Human Cells Identify Host Factors Used by Pathogens
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DOI：
10.1126/science.1178955
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发表时间：
2009-11-27
期刊：
影响因子：
56.9
通讯作者：
Brummelkamp, Thijn R.
中科院分区：
文献类型：
--
作者：
Carette, Jan E.;Guimaraes, Carla P.;Brummelkamp, Thijn R.
模型生物中功能丧失的遗传筛选已经阐明了许多生物学过程，但是哺乳动物细胞的二倍体基因组排除了大规模基因破坏。我们使用插入性诱变来开发一种筛选方法，以生成除8染色体以外的所有染色体的人类细胞系单倍体中的无效等位基因。使用这种方法，我们确定了宿主因子因流感和基因而编码的宿主因子，这些宿主因素和基因编码的重要元素，这些宿主因子途径的重要元素是生物合成途径的重要元素这是白喉毒素和外毒素A的细胞毒性作用所必需的。我们还鉴定了细胞骨化静态毒素作用所需的基因，包括与毒素相互作用的细胞表面蛋白。这种方法与单倍体酵母中的遗传方法具有概念和实用性。
Loss-of-function genetic screens in model organisms have elucidated numerous biological processes, but the diploid genome of mammalian cells has precluded large-scale gene disruption. We used insertional mutagenesis to develop a screening method to generate null alleles in a human cell line haploid for all chromosomes except chromosome 8. Using this approach, we identified host factors essential for infection with influenza and genes encoding important elements of the biosynthetic pathway of diphthamide, which are required for the cytotoxic effects of diphtheria toxin and exotoxin A. We also identified genes needed for the action of cytolethal distending toxin, including a cell-surface protein that interacts with the toxin. This approach has both conceptual and practical parallels with genetic approaches in haploid yeast.