Transgene Expression in the Retina

视网膜中的转基因表达

• 批准号: 7014004
• 负责人: GARY G MATTHEWS
• 金额: $ 14.7万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7014004
• 关键词: artificial chromosomes; biotechnology; confocal scanning microscopy; gene expression; gene targeting; genetic models; genetic regulatory element; genetically modified animals; green fluorescent proteins; model design /development; neurogenetics; neurons; plasmids; protein engineering; retina; technology /technique development; transfection; transfection /expression vector; zebrafish

DESCRIPTION (provided by applicant): The goal of this project is to develop techniques for the transgenic expression of altered proteins in retinal neurons. These techniques will be important in establishing the functional role of individual proteins in the retina. With the rapid success of genome sequencing projects, there is increasing information about gene sequences but little information about the functional roles of the proteins encoded by those genes. Transgenic techniques are particularly suitable for the task of deciphering those functional roles. Targeted expression of altered proteins or peptide fragments in particular cell types will allow the physiological action of a protein of interest to be tested in an individual cell. Also, the incorporation of a transgene in which expression of a marker protein is controlled by a target gene's regulatory region can be used to determine the cell types that express a particular gene. To accomplish these goals, methods will be developed that will allow the expression of engineered proteins in retinal neurons. Both conventional bacterial plasmid vectors and bacterial artificial chromosomes (BACs) will be explored for producing DNA constructs that are suitable for transgenesis. The transgenic techniques that are the goal of this project can be combined with electrophysiological and imaging techniques to yield a powerful way of attacking fundamental questions of retinal neurobiology. More broadly, completion of the proposed project will lead to new experimental tools that will be generally useful to retina researchers for the next level of functional analysis, after the completion of genome sequencing initiatives.

描述（由申请人提供）：本项目的目标是开发视网膜神经元中改变蛋白质的转基因表达技术。这些技术将是重要的，在建立视网膜中的单个蛋白质的功能作用。随着基因组测序计划的迅速成功，关于基因序列的信息越来越多，但关于这些基因编码的蛋白质的功能作用的信息却很少。 转基因技术特别适合于破译这些功能角色的任务。改变的蛋白质或肽片段在特定细胞类型中的靶向表达将允许在单个细胞中测试感兴趣的蛋白质的生理作用。此外，其中标记蛋白的表达受靶基因的调节区控制的转基因的掺入可用于确定表达特定基因的细胞类型。为了实现这些目标，将开发允许在视网膜神经元中表达工程蛋白的方法。常规细菌质粒载体和细菌人工染色体（BAC）都将被探索用于产生合适的DNA构建体， 转基因本项目的目标是将转基因技术与电生理学和成像技术相结合，以产生一种强有力的方法来解决视网膜神经生物学的基本问题。更广泛地说，完成拟议的项目将导致新的实验工具，这将是普遍有用的视网膜研究人员的下一个层次的功能分析，完成后的基因组测序计划。