Transgene Expression in the Retina

描述(由申请人提供)：该项目的目标是开发视网膜神经元中改变蛋白的转基因表达技术。这些技术对于确定单个蛋白质在视网膜中的功能作用将是重要的。随着基因组测序项目的迅速成功，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。针对特定细胞类型的改变的蛋白质或多肽片段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，标记蛋白的表达由目标基因的调节区控制的转基因的加入也可以用来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。传统的细菌质粒载体和细菌人工染色体(BAC)都将被用来产生合适的dna结构。 用于转基因。转基因技术是该项目的目标，可以与电生理和成像技术相结合，产生一种有效的方法来解决视网膜神经生物学的基本问题。更广泛地说，拟议项目的完成将带来新的实验工具，这些工具将在基因组测序计划完成后，对视网膜研究人员进行下一阶段的功能分析普遍有用。

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.