Role of the Lens1/Foxe3 Gene Family in Lens Formation

Lens1/Foxe3 基因家族在晶状体形成中的作用

• 批准号: 7072162
• 负责人: MILAN Alexander JAMRICH
• 金额: $ 33.07万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7072162
• 关键词: Xenopus oocyte; anterior chamber; clinical research; developmental genetics; eye disorder; gene mutation; genetically modified animals; histogenesis; human subject; in situ hybridization; laboratory mouse; lens; vertebrate embryology

DESCRIPTION (provided by applicant): Lens formation requires the interaction of several signaling molecules and transcriptional regulators. One of them is the murine forkhead protein Foxe3 and its Xenopus functional homologue Xlens1. We have shown previously that Foxe3 is expressed during the earliest stages of lens formation, and mutations in Foxe3 are the cause of the dysgenetic lens (dyl) phenotype in mouse. Mutations in human FOXE3 can cause anterior segment dysgenesis and cataracts. Overexpression of Xlens1 interferes with normal differentiation of lens fiber cells and results in overproliferation of cell in the anterior lens epithelium. The goal of this research is to identify the function and regulatory elements of Foxe3/Lens1 gene family. We propose the following specific aims related to lens formation. Specific Aim 1: Characterization of proteins that are involved in regulation of Foxe3 transcription. The goal of this specific aim is to isolate proteins that bind to the Foxe3 regulatory region and therefore are likely to be direct regulators of Foxe3 transcription. Specific Aim 2. Analysis of lens development in the absence of Foxe3/Xlens 1 function. The effects of elimination of Foxe3 function on development of the lens and on lens specific gene expression will be monitored in Foxe3 "knockout" mice. Elimination of Xlens1 function will be achieved by injection of Xenopus embryos with morpholinos directed against the translation initiation region of Xlens1. These experiments will determine the consequences of elimination of Xlens1 activity on development of structures derived from the anterior placodal region. Specific Aim 3. Correction of molecular and phenotypic defects in dysgenetic lens mutant mice by intrauterine gene transfer. In order to achieve this goal, we will introduce the wild type Foxe3 gene into dysgenetic lens embryos using viral vectors. These vectors will carry the wild type Foxe3 regulatory and coding sequences and will be delivered to the mutant embryos via intrauterine gene transfer.

描述（由申请人提供）：晶状体的形成需要多种信号分子和转录调节因子的相互作用。其中之一是小鼠叉头蛋白 Foxe3 及其非洲爪蟾功能同源物 Xlens1。我们之前已经证明 Foxe3 在晶状体形成的最早阶段表达，Foxe3 的突变是小鼠晶状体发育不良 (dyl) 表型的原因。人类 FOXE3 突变可导致眼前节发育不全和白内障。 Xlens1 的过度表达会干扰晶状体纤维细胞的正常分化，并导致前晶状体上皮细胞过度增殖。本研究的目的是鉴定 Foxe3/Lens1 基因家族的功能和调控元件。我们提出以下与晶状体形成相关的具体目标。 具体目标 1：表征参与 Foxe3 转录调节的蛋白质。这一具体目标的目的是分离与 Foxe3 调控区结合的蛋白质，因此可能是 Foxe3 转录的直接调控因子。 具体目标 2. 在没有 Foxe3/Xlens 1 功能的情况下分析镜片的开发。将在Foxe3“敲除”小鼠中监测消除Foxe3功能对晶状体发育和晶状体特异性基因表达的影响。通过向非洲爪蟾胚胎注射针对 Xlens1 翻译起始区的吗啡啉，可消除 Xlens1 功能。这些实验将确定消除 Xlens1 活性对源自前基板区域的结构发育的影响。 具体目标 3. 通过宫内基因转移纠正发育不良晶状体突变小鼠的分子和表型缺陷。为了实现这一目标，我们将使用病毒载体将野生型 Foxe3 基因引入发育不良的晶状体胚胎中。这些载体将携带野生型 Foxe3 调控和编码序列，并将通过宫内基因转移传递至突变胚胎。