REGULATION OF DNA EXPRESSION PLASMIDS IN TONGUE

舌头中 DNA 表达质粒的调控

• 批准号: 2132963
• 负责人: Stephen L-K Yen
• 金额: $ 4.11万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 1997-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2132963
• 关键词: gene expression; gene induction /repression; gene therapy; genetic promoter element; laboratory mouse; method development; plasmids; reporter genes; tongue; transfection

The delivery of an exogenous genetic element to cells and tissues in vivo has the potential to alter either their phenotype or pattern of differentiation if the DNA is transcribed and translated to produce a functional protein. In addition if this protein is secreted from the cells its circulation throughout the organism could result in a systemic effect due to the newly introduced DNA. Several different techniques have been developed to introduce new genetic sequences into cells, each having a particular set of strengths and weaknesses. procedures have been developed in our laboratory to inject naked plasmid DNA directly into tongue muscle such that the expression of the gene encoded by the DNA occurs. The tongue muscle presents several advantages for the delivery of new genes; including ready accessibility, non-proliferative cell population, high vascular flow, and uniform dense striated muscle, which have been used to advantage in the techniques developed in our laboratory. The method that has been developed has resulted in expression of reporter genes at the site of injection, and delivery of expressed proteins into the circulation for a systemic effect. The present studies are proposed to further develop this system and determine the applicability of inducible promoters and the time course for gene expression. The hypothesis for this project is; DNA expression plasmids directly injected into tongue muscle can be regulated through the use of inducible promoters. This hypothesis will be tested with two specific aims; 1) To determine whether inducible promoters included in a DNA expression plasmid will permit control of the temporal and quantitative patterns of expression of the reporter gene following injection into tongue muscle, 2) To determine whether the injected DNA plasmids will persist in the tongue muscle for extended periods and permit periodic or delayed induction and subsequent expression of the reporter gene. The longterm goal of this research is to develop the tongue injection model as a system suitable for gene therapy.

将外源遗传元件递送至体内细胞和组织 有可能改变它们的表型或模式 如果 DNA 被转录和翻译以产生分化 功能性蛋白质。 另外如果这种蛋白质是从细胞中分泌出来的 它在整个生物体中的循环可能会导致全身效应 由于新引入的DNA。 已经采用了几种不同的技术 开发用于将新的基因序列引入细胞，每个基因序列都有一个 特定的一组优点和缺点。 程序已经制定 在我们的实验室将裸露的质粒 DNA 直接注射到舌肌中 从而发生由DNA编码的基因的表达。 舌头 肌肉在传递新基因方面具有多种优势；包括 易于接近，非增殖细胞群，高血管流量， 和均匀致密的横纹肌，这在 我们实验室开发的技术。 已经开发出的方法 导致报告基因在注射部位表达，并且 将表达的蛋白质输送到循环中以产生全身作用。 目前的研究旨在进一步开发该系统 确定诱导型启动子的适用性和时间进程 基因表达。 该项目的假设是； DNA表达 直接注射到舌肌中的质粒可以通过 使用诱导型启动子。 这个假设将用两个测试来检验 具体目标； 1) 确定诱导型启动子是否包含在 DNA 表达质粒将允许控制时间和数量 注射到舌头后报告基因的表达模式 肌肉，2) 确定注射的 DNA 质粒是否会持续存在 舌头肌肉长时间工作并允许周期性或延迟 报告基因的诱导和随后的表达。 长期来看 这项研究的目标是将舌头注射模型开发为一个系统 适合基因治疗。