Analysis of Wilms tumour gene 1 (wt1) regulation and function in zebrafish

斑马鱼肾母细胞瘤基因 1 (wt1) 调控和功能分析

• 批准号: 28853747
• 负责人: Professor Dr. Christoph Englert
• 金额: --
• 依托单位: Leibniz-Institut für Alternsforschung - Fritz-Lipmann-Institut e.V. (FLI)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/28853747?language=en
• 关键词: Analysis; Wilms; tumour; gene; wt1

The tumour suppressor gene WT1/wt11 encodes a zinc finger transcription factor whose inactivation leads to Wilms tumour, a pediatric kidney cancer as well as to developmental anomalies concerning the urogenital system. Despite many years of research, mostly in man and mouse, knowledge about the precise physiologic and molecular function of the respective gene-product is scarce. We want to use the teleost fish Danio rerio (zebrafish) as a system to study the regulation and the physiologic role of wt1. The specific goals of this proposal are twofold. First, we want to know how wt1 expression is regulated in zebrafish. This will be analysed by the generation of GFP transgenic fish and subsequent biochemical experiments. The second goal is to elucidate the function of wt1 in development. We want to conduct overexpression and knockdown experiments using morpholinos. Finally, we want to identify molecules that mediate the activity of wt1. For this, knockdown of wt1 function and comparison of gene expression in normal and wt1-knockdown cells will be employed. Given the conservation of developmental processes across species we expect that the results which will be obtained through the experiments outlined here will contribute to the understanding of gene regulation, development and last but not least cancer biology also in higher vertebrates.

肿瘤抑制基因WT 1/wt 11编码锌指转录因子，其失活导致肾母细胞瘤，一种儿童肾癌以及泌尿生殖系统发育异常。尽管多年的研究，主要是在人和小鼠，知识的精确生理和分子功能的基因产物是稀缺的。本研究以斑马鱼（Danio rerio）为研究对象，研究wt 1基因的调控及其生理作用。这项建议的具体目标有两个方面。首先，我们想知道wt 1在斑马鱼中的表达是如何调节的。这将通过GFP转基因鱼的产生和随后的生化实验来分析。第二个目标是阐明wt 1在发育中的功能。我们想用吗啉代进行过表达和敲低实验。最后，我们想确定介导wt 1活性的分子。为此，将采用wt 1功能的敲低和正常和wt 1敲低细胞中基因表达的比较。考虑到物种间发育过程的保守性，我们期望通过本文概述的实验获得的结果将有助于理解高等脊椎动物的基因调控，发育以及最后但并非最不重要的癌症生物学。