Phenotypic screening using DNA-encoded libraries

使用 DNA 编码文库进行表型筛选

• 批准号: 10238888
• 负责人: Thomas J. Kodadek
• 金额: $ 59.51万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-04-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10238888
• 关键词: Affect; Agonist; Animals; Biochemical; Biological; Biological Assay; Biological Process; Biology; Categories; Cells; Charge; Cleaved cell; Complex; DNA; Development; Engineering; Environment; Exhibits; Fishes; Fluorescence; Generations; Goals; Incubated; Infrastructure; Laboratories; Libraries; Ligands; Link; Membrane; Molecular Target; Permeability; Pharmaceutical Preparations; Pharmacology; Phenotype; Population; Process; Proteins; RNA; Reporter Genes; Reporting; Research Personnel; Rest; Signal Transduction; Specialized Center; Structure; Surface; System; Technology; Tentagel resin; Whole Organism; Work; base; cellular engineering; chemotherapy; flasks; fluorescence activated cell sorter device; high throughput screening; innovation; interest; novel; novel strategies; screening; small molecule

Project Summary This project will focus on the development of a powerful new approach for the discovery of bioactive small molecules via cell-based phenotypic screening. DNA-encoded one bead one compound libraries built on small beads will be complexed with cells and suspended in solution. A photolabile linker will be cleaved, exposing the cells briefly to a high local concentration of the compound on the bead to which they are attached. The cells will be engineered to report a phenotype of interest through the increase of a fluorescent signal. Beads carrying positive cells will be isolated by FACS and the encoding tags sequenced, revealing the structures of the hits. Since we will use libraries of molecules capable of linking to targets covalently, the identification of the target will be simplified greatly. This system will be used to discover novel new chemotherapy agents.

项目摘要 该项目将重点开发一种强大的新方法，用于发现生物活性小分子。 分子通过基于细胞的表型筛选。DNA编码的一珠一化合物文库构建在小 珠粒将与细胞复合并悬浮在溶液中。光不稳定的接头将被切割，暴露出 使细胞短暂地接触到它们所附着的珠粒上的高局部浓度的化合物。细胞 将被工程化以通过荧光信号的增加报告感兴趣的表型。珠携带 通过FACS分离阳性细胞，并对编码标签进行测序，揭示命中物的结构。 由于我们将使用能够与靶共价连接的分子库，因此靶的鉴定 将大大简化。该系统将用于发现新的化疗药物。