Characterizing a novel promoter of mouse MOR-1 gene

描述（由申请人提供）：本申请的主要目标 是为了更好地了解转录调控的亩 阿片受体基因（Oprm）， 技术.克隆莫尔-1 cDNA后不久，启动子（El启动子） 和结构（外显子1-4）的原始莫尔-1基因进行了鉴定， 表征了随后很快分离出两种剪接形式， 莫尔-1A和MOR-1B。M0 R-1基因的多样性进一步通过以下说明： 我们最近发现了另外9个外显子，12个剪接变异体， 以及与其中一个新外显子相关的新启动子（E11启动子）。这 应用程序的重点是阐明详细的结构和功能， E11启动子及其与E1启动子的关系。E11启动子是 位于原E1启动子上游约10 kb处。替代 由E11启动子驱动的转录本剪接产生八种变体 编码许多新蛋白以及原始的莫尔-1蛋白。 由E11启动子调控的三种变体产生原始的莫尔-1 蛋白因此，这种单一蛋白质可以由四种剪接变体产生， μ阿片受体基因在两个不同的启动子的控制下。 不同脑区的差异表达揭示了 区域特异性RNA加工。具有核心的神经元启动子活性 启动子和负元件。具体目标是 本研究的主要目的是：（1）鉴定E11顺式作用元件 启动子区突变分析;（2）DNA结合蛋白的分离 （3）E11启动子的研究， 在转基因小鼠中与E1启动子的关系。我的长期目标是 了解莫尔-1基因的调控机制， 深入了解其药理学和生理学意义， 调控E11启动子及其相关变体的意义在于 这是由反义定位研究提出的，其中两个反义探针靶向 外显子11阻断脊髓吗啡和脊髓上M6 G镇痛。知识 从应用程序中获得的信息将有助于确定复杂性， MOR-1基因调控的功能重要性，建立细胞模型， 为研究其基因调控提供转基因动物模型， 用于开发用于疼痛控制和药物治疗的新药物的潜在靶点 虐待

DESCRIPTION (provided by the applicant): The primary goal of this application is to obtain a better understanding of transcriptional regulation of the mu opioid receptor gene (Oprm) through molecular biological means and transgenic techniques. Soon after the MOR-1 cDNAs were cloned, the promoter (El promoter) and structure (exons 1-4) of the original MOR-1 gene were identified and characterized. This was soon followed by the isolation of two splice forms, MOR-1A and MOR-IB. The diversity of the MOR-l gene was further illustrated by our recent identification of an additional nine exons, twelve splice variants and a novel promoter associated with one of the new exons (E11promoter). This application focuses on elucidating the detailed structure and function of the E11 promoter and its relationships with E1 promoter. The E11 promoter is located at about10 kb upstream of the original E1 promoter. Alternative splicing of the transcripts driven by the E11 promoter yield eight variants encoding a number of novel proteins as well as the original MOR-1 protein. Three of the variants regulated by the E11 promoter generate the original MOR-1 protein. Thus, this single protein can be generated by four splice variants of the mu opioid receptor gene under the control of two distinct promoters. Differential expression of the variant mRNAs in various brain regions revealed region-specific RNA processing. Neuronal promoter activities with a core promoter and a negative element have been suggested. The specific aims to achieve the overall goal are: (1) Identification of cis-acting elements in E11 promoter region through mutant analysis; (2) Isolation of DNA-binding proteins that regulate promoter activities; (3) Investigation of the E11 promoter and its relationships with E1 promoter in transgenic mice. My long-term goal is to understand the mechanisms by which the MOR-1 gene is regulated and to gain insights into the pharmacological and physiological significance of its regulation. The significance of the E11 promoter and its associated variants is suggested by antisense mapping studies in which two antisense probes targeting exon 11 blocked spinal morphine and supraspinal M6G analgesia. The knowledge obtained from the application will help to determine the complexity and functional importance of MOR-l gene regulation, establish the cell models and the transgenic animal models for studying its gene regulation, and provide potential targets for developing novel drugs useful in pain control and drug abuse.