Novel Single Cell Reporter Gene Assay

新型单细胞报告基因检测

• 批准号: 6878265
• 负责人: YEVGENYA AKSELBAND
• 金额: $ 16.64万
• 依托单位: ONE CELL SYSTEMS, INC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6878265
• 关键词: Novel; Single; Cell; Reporter; Gene

DESCRIPTION (provided by applicant): This Small Business Innovation Research Phase I project aims to develop a novel reporter gene assay for studying eukaryotic gene expression and regulation in individual transfected cells, and for isolating variants with defined transcription responses for cloning and use in subsequent studies. This assay will use single cell gel microdrop technology, a secreted enzyme capture and detection format, and fluorescence activated cell sorting. Although there have been significant developments in reporter gene assays, an assay which provides high sensitivity, high throughput and single cell resolution is still needed. Frequently, gene products may be difficult to detect due to low abundance or instability. In addition, many potential therapeutic molecules that modulate transcription, exhibit weak target binding affinities, resulting in low reporter gene expression. The proposed assay will permit detection of a wide range of signal intensities from single, transfected cells which can be isolated for subsequent cloning. Any study of a biochemical process that terminates in a transcriptional event would benefit from use of the proposed reporter gene assay. This assay will facilitate drug discovery by permitting rapid screening of combinatorial chemistry or natural product libraries.

描述（由申请人提供）：这个小型企业创新研究阶段项目旨在开发一种新的记者基因测定，用于研究单个转染的细胞中的真核基因表达和调节，并用于隔离具有定义的转录响应的变体以在后续研究中进行克隆和使用。该测定法将使用单细胞凝胶微旋转技术，一种分泌的酶捕获和检测格式以及荧光激活的细胞分选。尽管在报告基因测定方面已经有很大的发展，但仍需要高灵敏度，高吞吐量和单细胞分辨率的测定法。通常，由于丰度低或不稳定性，基因产物可能难以检测。此外，许多调节转录的潜在治疗分子表现出弱靶结合亲和力，导致报告基因表达低。所提出的测定将允许检测来自单个，转染的细胞的广泛信号强度，这些信号强度可以分离以进行后续克隆。任何在转录事件中终止的生化过程的研究都将受益于使用拟议的记者基因测定法。该测定法可以通过允许快速筛选组合化学或天然产品库来促进药物发现。