YEAST BASED NUCLEAR REPORTER LIFETIME FRET ASSAY SYSTEM

我们将开发一个由仪器、试剂和 高通量全细胞核报告基因快速检测方法 药物发现应用中先导化合物的筛选(HTS)。这个 这种方法是基于使用绿色荧光蛋白作为 细胞内市场，以及荧光寿命传感来量化 报告基因的表达。该方法的一个关键创新是使用 最近开发的一种新的荧光寿命传感技术 由Ciencia提供，可在坚固、低成本、紧凑的环境中实施 仪器，并且能够高灵敏度、高寿命 分辨率高，测量速度快。提出的寿命感测 该方法将解决当前基于强度的一些基本问题 荧光系统，如因光漂白而产生的伪影和非 特定的细胞自发荧光背景和有色干扰 样品基质中的化合物。我们的长期目标是发展 紧凑、坚固、灵敏、低成本的仪器和分析 药物发现中的吞吐量筛选。拟议的工作解决了 面临竞争压力的制药公司的需求 将成功的治疗新产品更快更多地推向市场 效率很高。据我们所知，这将是第一次 利用荧光寿命传感进行高通量筛选 申请。 建议的商业应用： 在铅药物发现、细胞和分子方面的高通量筛选 生物研究仪器、临床诊断、农业、 食品加工，环境分析取证。

We will develop a system consisting of instrumentation, reagents and methods for rapid whole cell nuclear reporter assays for high throughput screening (HTS) of lead compounds in drug discovery applications. The approach is based on the use of green fluorescent proteins as an intracellular market, and fluorescence lifetime sensing to quantify reporter gene expression. A key innovation of the approach is the use of a new fluorescence lifetime sensing technology, recently developed by Ciencia which can be implemented in rugged, low-cost, compact instrumentation, and which is capable of high sensitivity, high lifetime resolution, and rapid measurement time. The proposed lifetime sensing approach will solve some fundamental problems of current intensity-based fluorescence system, such as artifacts due to photobleaching and non- specific cell autofluorescence background, and interference from colored compounds in the sample matrix. the long-term objective is to develop compact, robust, sensitive, low-cost instrumentation and assays for high throughput screening in drug discovery. the proposed work addresses the needs of pharmaceutical companies which are under competitive pressures to bring successful new therapeutic products to market faster and more efficiently. To our knowledge, this would be the first time that fluorescence lifetime sensing is exploited for high throughput screening applications. PROPOSED COMMERCIAL APPLICATION: High throughput screening in lead drug discovery, cell and molecular biology research instrumentation, clinical diagnostics, agriculture, food processing , environmental analysis forensics.