OTHER FUNCTIONS SBIR PHASE II TOPIC 290 "RNAI SYNTHETIC LETHAL SCREEN IN CANCER

抑制合成致死(SL)(协同)DNA损伤和修复(DDR)药物靶点功能的可能性是开发高特异性和低毒性抗癌药物的一个有前景的方向。正如290份SBIR合同提案中概述的那样，识别SL靶标的一个有希望的、但尚未实现的战略是，基于从一套完整的DDR基因中去除两个靶标，应用系统的、无偏见的RNAi活性筛查。作为构建癌症-SL基因相互作用图谱的第一步，使用第一阶段SBIR合同资金，我们开发了构建新的双特异池慢病毒shRNA文库的技术，该文库旨在组合击倒一组合理选择的基因中的任何两个靶点。此外，我们还成功地应用双特异性shRNA文库检测了40个DDR基因在等基因乳腺癌细胞模型中的SL二元相互作用。第二阶段合同研究的最终科学目标是将开发的组合RNAi筛选技术应用于大规模发现来自三种癌细胞系的400个DDR靶点中所有可能的SL基因对。作为拟议研究的实际成果，我们将开发、验证和商业化一个新的开源SL DDR RNAi筛选平台，用于无偏见地发现和注释癌细胞模型中SL相互作用。为了方便研究人员使用组合RNAi筛选技术，Cellecta将在癌细胞模型中提供定制的双特异性shRNA文库和定制的RNAi筛选服务。

A promising direction in the development of anti-cancer drugs with high specificity and low toxicity is the possibility of inhibiting the functions of synthetically lethal (SL) (synergetic) DNA Damage and Repair (DDR) drug targets that are specifically modulated in cancer development. As outlined in the 290 SBIR contract proposals, one of the promising, although unrealized, strategies for identifying SL targets is to apply a systematic, unbiased RNAi viability screen based on the ablation of two targets from a complete set of DDR genes. As a first step for constructing a cancer-SL gene interaction map, which used Phase I SBIR contract funding, we developed the technology for the construction of novel bi-specific pooled lentiviral shRNA libraries designed for the combinatorial knockdown of any two targets in a set of rationally selected genes. Furthermore, we successfully demonstrated the application of the bi-specific shRNA library to detect SL binary interactions between 40 DDR genes in an isogenic breast cancer cell model. The ultimate scientific goal of the Phase II contract studies is to apply the developed combinatorial RNAi screening technology to the large-scale discovery of all possible SL gene pairs in a set of 400 DDR targets from a panel of three cancer cell lines. As a practical outcome of the proposed studies, we will develop, validate and commercialize a novel open-resource SL DDR RNAi screening platform for the unbiased discovery and annotation of SL interactions in cancer cell models. To provide researchers with easy access to the combinatorial RNAi screening technology, Cellecta will offer custom bi-specific shRNA libraries and custom RNAi screening services in cancer cell models.