A multiplex genome-wide shRNA screening platform for cancer-lethal gene discovery

描述(申请人提供)：设计有效的癌症治疗靶向疗法的一个主要瓶颈是确定相关的药物靶点。基因筛查在基因组规模上衡量癌细胞和正常细胞中单个基因抑制的结果，是可以识别这种药物靶点的强大实验。直到最近，这些筛查还不能在人类细胞中进行。为了进行这样的筛选，我的实验室开发了基于RNA干扰(RNAi)原理的新方法，以单独灭活人类基因组中的每一个基因，大约32,000个基因，一次一个。此外，我们还开发了以高通量方式进行这些筛查的技术。通过这次融资机会，我的目标是进一步发展我们的RNAi技术平台，以提高其吞吐量和保真度。我将使用肺癌细胞系作为这项技术开发工作的发现范例。最终目标是实现对大量癌细胞系的全基因组询问，以全面识别癌细胞特有的依赖性和脆弱性，而不是正常细胞。这种方法应该会发现许多以前未被认识到的药物发现目标，并有可能为癌症患者提供更多、更有效的治疗选择。

DESCRIPTION (provided by applicant): A major bottleneck in devising effective targeted therapies for cancer treatment lies at the identification of relevant drug targets. Genetic screens that measure the outcome of inhibiting individual genes in cancer cells and in normal cells on a genome-scale are powerful experiments that could identify such drug targets. Until recently these screens could not be performed in human cells. To perform such screens, my laboratory has developed new methods, based on the principle of RNA interference (RNAi), to individually inactivate every gene in the human genome, approximately 32,000 genes, one at a time. Furthermore, we have developed technologies to carry out these screens in high throughput fashion. Through this funding opportunity, I aim to further develop our RNAi technology platform to enhance its throughput and fidelity. I will use lung cancer cell lines as a discovery paradigm for this technology development effort. The ultimate goal is to enable the genome-wide interrogation of large numbers of cancer cell lines to comprehensively identify the dependencies and vulnerabilities specific to cancer cells but not normal cells. This approach should uncover many previously unrecognized targets for drug discovery and has the potential to provide additional, more effective treatment options for cancer patients.