SBIR Phase I: A Deep Sequencing-Based Platform for Yeast Two-Hybrid Screening

这个小企业创新研究第一阶段项目涉及应用Illumina的Solexa深度测序技术作为酵母双杂交（Y2H）筛选的读出。提出的研究源于认识到Y2H筛选的主要限制是缺乏筛选结果的真正大规模读数。应用Illumina的深度测序方法来全面分析汇集的Y2H菌株，将有效地取代传统方法所需的单个克隆的时间和劳动密集型测序。此外，量身定制的评分方案将允许使用定量统计分析来确定Y2H读数的重要性。因此，深度测序技术的这种特殊增强不仅将大大提高Y2H筛选的吞吐量（1,000-10,000倍），而且还将为鉴定真正的阳性结果提供高可信度。这项研究的更广泛/商业影响将是使用Y2H和相关程序进行筛选的强有力转变。一旦这种Y2H筛选技术建立起来，研究人员将能够使用转录组分析的工具和工作流程，识别和阐明蛋白质-蛋白质相互作用的复杂网络，这些网络基本上是所有病理和复杂疾病的基础。商业应用将针对那些希望探索药物靶点和致癌基因的生物学背景的学术和工业实验室。与传统的Y2H方法相比，该方法将以更低的成本提供更好的结果。虽然Quintara在基因组学应用方面的商业机会非常大，但真正的好处将会给研究界和人类健康带来好处。

This Small Business Innovation Research Phase I project concerns applying Illumina's Solexa deep-sequencing technology as a read-out for yeast two-hybrid (Y2H) screens. The proposed research arose from the recognition that the principal limitation of Y2H screening is the lack of a true large-scale readout for the screening results. Applying Illumina's deep-sequencing methodology to comprehensively analyze pooled Y2H strains will effectively supplant the time- and labor-intensive sequencing of individual clones required by conventional approaches. Further, a tailored scoring scheme will allow use of quantitative statistical analysis to determine the significance of the Y2H readouts. Hence, this particular enhancement of deep-sequencing technology will not only vastly increase the throughput of Y2H screens (1,000-10,000 fold), but will also provide high confidence in identification of true positive results.The broader/commercial impact of this research will be a strong shift towards using Y2H and related procedures for screening. Once this Y2H screening technology is established, researchers, using tools and workflows set up for transcriptome analysis, will be able to identify and elucidate the intricate networks of protein-protein interactions that underlie essentially all pathologies and complex diseases. Commercial applications will target academic and industrial labs that wish to explore the biological context of drug targets and oncogenes. Compared with conventional Y2H methods, the proposed approach will deliver superior results at a much lower cost. While the business opportunity for Quintara for genomics applications is very large, the real benefits will accrue to the research community and human health.