DESIGN OF TARGETED SYNTHETIC GENE DELIVERY VEHICLES

靶向合成基因递送载体的设计

• 批准号: 2862085
• 负责人: ILYA V KOLTOVER
• 金额: $ 3.03万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-07 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2862085
• 关键词: bioengineering /biomedical engineering; biological signal transduction; biophysics; cell population study; chemical conjugate; chemical stability; chemical structure; confocal scanning microscopy; drug delivery systems; gene expression; gene therapy; genetic transcription; green fluorescent proteins; intermolecular interaction; polymers; tissue /cell culture; tissue engineering; transfection /expression vector

We propose a set of fundamental studies aimed at understanding the mechanistic basis for improving efficiency of gene delivery to cells via targeted molecular conjugate (polyplex) vectors. We plan to isolate the expression-limiting steps involved in the overall process of transgene expression: cell surface binding, internalization, endosomal escape, nuclear localization and transcription, and to elucidate the molecular properties of the conjugate governing each step. This goal will be achieved by constructing a modular conjugate, permitting selective modification of its molecular properties. GFP will serve as the transgene, allowing quantitative measurement of both expression efficiency and level across a cell population distribution. Our battery of experimental approaches includes quantitative assays for binding and internalization, confocal and two-photon microscopy visualization of conjugate localization within cells, quantitative measurements of expression levels and detailed physico-chemical analyses of conjugate structure and stability.

我们提出了一套基础研究，旨在了解通过靶向分子偶联物（复合物）载体提高基因递送到细胞的效率的机制基础。 我们计划分离的表达限制的步骤，在转基因表达的整个过程中：细胞表面结合，内化，内体逃逸，核定位和转录，并阐明每个步骤的共轭物的分子特性。 这一目标将通过构建模块化缀合物来实现，从而允许选择性地修饰其分子性质。 GFP将作为转基因，允许跨细胞群体分布的表达效率和水平的定量测量。 我们的实验方法的电池包括定量测定结合和内化，共聚焦和双光子显微镜可视化共轭定位在细胞内，定量测量的表达水平和详细的物理化学分析共轭结构和稳定性。