High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities
High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities
复制标题
DOI：
10.1016/j.cell.2015.11.015
复制
发表时间：
2015-12-06
期刊：
影响因子：
64.5
通讯作者：
Moffat, Jason
中科院分区：
文献类型：
--
作者：
Hart, Traver;Chandrashekhar, Megha;Moffat, Jason
扰乱人类细胞中基因的能力对于阐明基因功能至关重要，并且在寻找癌症等疾病的治疗靶点方面具有巨大的潜力。为了扩大人类核心和环境相关适应度基因的目录，我们开发了一个高复杂性的第二代基因组级CRISPR-Cas9 gRNA文库，并将其应用于五种人类细胞系的适应度筛选。使用改进的贝叶斯分析方法，我们不断发现比以前观察到的多5倍的适合基因。我们提出了1,580个人类核心适应性基因的列表，并描述了它们的一般特性。此外，我们证明了上下文依赖的适应度基因准确地概括了由已知癌基因诱导的途径特异性遗传脆弱性，并揭示了对特定受体酪氨酸激酶的细胞类型特异性依赖，即使在致癌KRAS背景下也是如此。因此，使用高复杂性CRISPR-Cas9文库对人类细胞系适应度基因进行严格鉴定，可以提供细胞遗传脆弱性的高分辨率视图。
The ability to perturb genes in human cells is crucial for elucidating gene function and holds great potential for finding therapeutic targets for diseases such as cancer. To extend the catalog of human core and context-dependent fitness genes, we have developed a high-complexity second-generation genome-scale CRISPR-Cas9 gRNA library and applied it to fitness screens in five human cell lines. Using an improved Bayesian analytical approach, we consistently discover 5-fold more fitness genes than were previously observed. We present a list of 1,580 human core fitness genes and describe their general properties. Moreover, we demonstrate that context-dependent fitness genes accurately recapitulate pathway-specific genetic vulnerabilities induced by known oncogenes and reveal cell-type-specific dependencies for specific receptor tyrosine kinases, even in oncogenic KRAS backgrounds. Thus, rigorous identification of human cell line fitness genes using a high-complexity CRISPR-Cas9 library affords a high-resolution view of the genetic vulnerabilities of a cell.