A Genome-wide CRISPR Death Screen Identifies Genes Essential for Oxidative Phosphorylation.
A Genome-wide CRISPR Death Screen Identifies Genes Essential for Oxidative Phosphorylation.
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DOI：
10.1016/j.cmet.2016.08.017
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发表时间：
2016-12-13
期刊：
影响因子：
29
通讯作者：
Mootha VK
中科院分区：
文献类型：
--
作者：
Arroyo JD;Jourdain AA;Calvo SE;Ballarano CA;Doench JG;Root DE;Mootha VK
氧化磷酸化(OXPHOS)是人类产生三磷酸腺苷的主要途径。OXPHOS的缺陷可由线粒体或核基因组的突变引起，并构成最大的先天性新陈代谢错误集合。目前，我们缺乏OXPHOS所必需的人类基因和途径的完整目录。在这里，我们介绍了一个全基因组的CRISPR“死亡屏幕”，它主动选择死亡的细胞，以揭示OXPHOS所需的人类基因，灵感来自于经典的观察结果，即缺乏OXPHOS的人类细胞在葡萄糖中生存，但在半乳糖中死亡。我们报告了191个对OXPHOS必不可少的高置信度命中，包括72个潜在的已知OXPHOS疾病。我们的屏幕显示了一个由NGRN、WBSCR16、RPUSD3、RPUSD4、TRUB2和FASTKD2组成的功能模块，它调节线粒体16S rRNA和线粒体内翻译。我们的工作产生了OXPHOS所需的丰富的基因目录，更广泛地说，展示了死亡筛查在功能基因组分析中的力量。
Oxidative phosphorylation (OXPHOS) is the major pathway for ATP production in humans. Deficiencies in OXPHOS can arise from mutations in either mitochondrial or nuclear genomes and comprise the largest collection of inborn errors of metabolism. At present we lack a complete catalog of human genes and pathways essential for OXPHOS. Here we introduce a genome-wide CRISPR “death screen” that actively selects dying cells to reveal human genes required for OXPHOS, inspired by the classic observation that human cells deficient in OXPHOS survive in glucose but die in galactose. We report 191 high-confidence hits essential for OXPHOS, including 72 underlying known OXPHOS diseases. Our screen reveals a functional module consisting of NGRN, WBSCR16, RPUSD3, RPUSD4, TRUB2, and FASTKD2 that regulates the mitochondrial 16S rRNA and intra-mitochondrial translation. Our work yields a rich catalog of genes required for OXPHOS and, more generally, demonstrates the power of death screening for functional genomic analysis.