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

该项目的长期目标是更好地了解 调节牙齿发育的遗传过程，最终致力于 基因治疗和牙齿置换。该项目的重点是转录 上调的丝氨酸蛋白酶EMSP1编码基因的调控 在釉质发生的成熟期。假设EMSP1是 以高度受限的、特定于组织的方式表达，并且通过定义 参与调控EMSP1表达的分子信号，洞察力将 进入监管级联，导致从秘密到 成釉作用的成熟期。三个具体目标是：(1)产生 并鉴定了一种β-半乳糖苷酶敲入小鼠，以准确地定义 EMSP1基因的时间和空间表达；(2)确定 小鼠EMSP1启动子；以及(3)鉴定小鼠EMSP1启动子 区域。敲门鼠将有一个报告基因转录自 内源性EMSP1启动子将被用来确定精确的时间 和EMSP1的空间表达。我们将把这种模式与 转基因小鼠和包括削减的转基因细胞系 启动子-报告者结构，以定义最小启动子 提供正常的表情。

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.