Genetic interaction screening in renal cell carcinoma

肾细胞癌的遗传相互作用筛查

• 批准号: 10926504
• 负责人: Michael Aregger
• 金额: $ 42.02万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10926504
• 关键词: CCR; Cell Line; Cells; Chromosome Mapping; Clustered Regularly Interspaced Short Palindromic Repeats; Dependence; Genes; Genetic; Genome; Goals; Libraries; Malignant Epithelial Cell; Metabolic; Metabolism; Patients; Regulation; Renal Cell Carcinoma; Somatic Mutation; Therapeutic; Translations; Validation; Western Blotting; combinatorial; fitness; functional genomics; genome-wide; insight; knockout gene; novel; nuclease; screening

We have obtained 6 patient-derived renal cell carcinoma (RCC) cell lines harboring somatic mutations in metabolic genes from the lab of Marston Linehan (UOB/CCR/NCI). We have transduced those RCC lines with CRISPR-Cas nucleases amenable to our CHyMErA combinatorial genome perturbation platform. Expression of Cas nucleases have been verified by western blotting and editing efficiency of the cell lines has been confirmed. Screens with genome-scale gene knockout libraries are currently in progress to identify fitness genes across the various environmental and genetic backgrounds. In a second project, we have generated a metabolism-focused genetic interaction library and are currently performing a pilot screen with this library.

我们从 Marston Linehan 实验室 (UOB/CCR/NCI) 获得了 6 个患者来源的肾细胞癌 (RCC) 细胞系，这些细胞系在代谢基因中存在体细胞突变。我们已经使用适合我们的 CHyMErA 组合基因组微扰平台的 CRISPR-Cas 核酸酶转导了这些 RCC 系。 Cas核酸酶的表达已通过蛋白质印迹验证，并且细胞系的编辑效率已得到证实。目前正在进行基因组规模基因敲除文库的筛选，以识别各种环境和遗传背景的适应性基因。在第二个项目中，我们生成了一个以代谢为中心的遗传相互作用库，目前正在使用该库进行试点筛选。