Genetic screens for target discovery in cancer

背景资料。使用RNAi和CRISPR/Cas9文库进行基因筛查是询问癌症基因组功能脆弱性的有效方法。我们已经开发了进行快速且经济有效的shRNA和sgRNA文库筛选的技术。目的。在这个项目中，我们的目标是利用RNAi和CRISPR/Cas9文库进行功能丧失和功能获得的基因筛查，以研究癌细胞的遗传脆弱性，并确定耐药机制。我们还将与其他研究人员(主要是NIH内壁计划的调查人员)合作，使他们能够在癌细胞系中进行筛查。重要的材料和方法。针对大多数已注释的人类基因的汇集shRNA和CRISP/Cas9文库。成就。我们已经在许多不同的癌症疾病领域完成了几项筛查。特别是，我们已经确定了可以调节KRAS突变癌细胞对MEK抑制剂敏感性的基因和遗传途径。我们正在研究来自这些筛选的候选基因。一份描述调查结果的手稿已经提交出版。

BACKGROUND. Genetic screen using RNAi and CRISPR/Cas9 libraries is a powerful approach to interrogate the cancer genome for functional vulnerabilities. We have developed technologies to conduct rapid and cost-effective shRNA and sgRNA library screens. PURPOSE. In this project we aim to carry out loss-of-function and gain-of-function genetic screens using RNAi and CRISPR/Cas9 libraries to study genetic vulnerabilities in cancer cells and identify mechanisms of drug resistance. We will also collaborate with other investigators (primarily investigators in the NIH Intramural Program) to enable them to conduct screens in cancer cell lines. SIGNIFICANT MATERIALS AND METHODS. Pooled shRNA and CRISP/Cas9 library targeting most annotated human genes. ACCOMPLISHMENT. We have completed several screens in a number of different cancer disease areas. In particular, we have identified genes and genetic pathways that could modulate KRAS mutant cancer cells' sensitivity to MEK inhibitors. We are studying candidate genes from these screens. A manuscript describing out findings have been submitted for publication.