Genetic screens in mammalian cells commonly focus on loss-of-function approaches. To evaluate the phenotypic consequences of extra gene copies, we used bulk segregant analysis (BSA) of radiation hybrid (RH) cells. We constructed six pools of RH cells, each consisting of ∼2500 independent clones, and placed the pools under selection in media with or without paclitaxel. Low pass sequencing identified 859 growth loci, 38 paclitaxel loci, 62 interaction loci, and three loci for mitochondrial abundance at genome-wide significance. Resolution was measured as ∼30 kb, close to single-gene. Divergent properties were displayed by the RH-BSA growth genes compared to those from loss-of-function screens, refuting the balance hypothesis. In addition, enhanced retention of human centromeres in the RH pools suggests a new approach to functional dissection of these chromosomal elements. Pooled analysis of RH cells showed high power and resolution and should be a useful addition to the mammalian genetic toolkit.

中文翻译：

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辐射杂交体的合并分析确定了哺乳动物细胞中生长和药物作用的位点。

哺乳动物细胞中的遗传筛选通常侧重于功能丧失方法。为了评估额外基因拷贝的表型后果，我们使用了辐射杂交 (RH) 细胞的批量分离分析 (BSA)。我们构建了六个 RH 细胞池，每个池由约 2500 个独立克隆组成，并将这些池置于有或无紫杉醇的培养基中进行选择。低通测序鉴定了 859 个生长位点、38 个紫杉醇位点、62 个相互作用位点和三个具有全基因组意义的线粒体丰度位点。分辨率测量为~30 kb，接近单基因。与功能缺失筛选的生长基因相比，RH-BSA 生长基因显示出不同的特性，驳斥了平衡假说。此外，人类着丝粒在 RH 池中的增强保留表明了一种新的方法来对这些染色体元素进行功能性解剖。RH 细胞的合并分析显示出高功率和分辨率，应该是对哺乳动物遗传工具包的有用补充。