REGULATION OF ENAMEL MATRIX SERINE PROTEINASE 1 (EMSP1) DURING AMELOGENESIS

本项目的长期目标是更好地了解 调节牙齿发育的遗传过程，最终 基因治疗和牙齿替换该项目的重点是转录 调节编码EMSP 1的基因，EMSP 1是一种丝氨酸蛋白酶， 在釉质形成的成熟阶段。假设EMSP 1是 以高度限制的组织特异性方式表达，并且通过定义 参与调节EMSP 1表达的分子信号， 进入调节级联反应，引起从分泌到 釉质形成的成熟阶段。三个具体目标是：（1）产生 并表征β-半乳糖苷酶敲入小鼠，以精确定义 EMSP 1基因的时间和空间表达;（2）定义EMSP 1基因的限制， 小鼠EMSP 1启动子;和（3）表征小鼠EMSP 1启动子 地区敲入小鼠将有一个报告基因转录自 内源性EMSP 1启动子，其将用于确定 和EMSP 1的空间表达。这一模式将与 转基因小鼠和转染的细胞系 启动子-报告基因构建体，以确定 给出了正常的表达。

The long term goal of this project is to better understand the genetic processes that regulate tooth development, to ultimately work toward gene therapy and tooth replacement. The project focuses on transcriptional regulation of the gene encoding EMSP1, which is a serine protease upregulated during the maturation stage of amelogenesis. The hypotheses are that EMSP1 is expressed in a highly restricted, tissue-specific manner, and that by defining the molecular signals involved in regulating EMSP1 expression, insight will be gained into the regulatory cascade causing the transition from secretory to maturation stages of amelogenesis. The three Specific Aims are: (1) To generate and characterize a beta-galactosidase knock-in mouse to precisely define the temporal and spatial expression of the EMSP1 gene; (2) to define the limits of the mouse EMSP1 promoter; and (3) to characterize the mouse EMSP1 promoter region. The knockin mouse will have a reporter gene transcribed from the endogenous EMSP1 promoter which will be used to determine the precise temporal and spatial expression of EMSP1. This pattern will be compared to that in transgenic mice and transfected cell lines that include cutback promoter-reporter constructs, in order to define the minimal promoter that gives normal expression.