REGULATION OF DNA EXPRESSION PLASMIDS IN TONGUE

将外源遗传因子传递到体内的细胞和组织 有可能改变他们的表型或模式， 如果DNA被转录和翻译以产生一种 功能蛋白 此外，如果这种蛋白质从细胞中分泌出来， 它在整个机体内的循环会导致全身效应 由于新引入的DNA。 已经有几种不同的技术 开发了将新的基因序列引入细胞，每个基因序列都有一个 特定的优势和劣势。 已经制定了程序， 将裸质粒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有几个优势，包括 容易接近，非增殖细胞群，高血管流量， 和均匀致密的横纹肌，这已被用来在优势， 我们实验室开发的技术。 已经开发的方法 导致报告基因在注射部位表达， 将表达的蛋白质递送到循环中以产生全身效应。 目前的研究建议进一步发展这一系统， 确定诱导型启动子的适用性和 基因表现。 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。 这一假设将用两个 具体目的：1）为了确定是否包括在诱导型启动子中， DNA表达质粒将允许控制时间和定量表达。 舌内注射后报告基因的表达模式 2）为了确定注射的DNA质粒是否会在肌肉中持续存在， 舌头肌肉延长的时期，并允许定期或延迟 报告基因的诱导和随后的表达。 的长期 本研究的目标是开发舌注射模型作为一个系统 适合基因治疗。

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.