Drosophila as a Model to Investigate Rett Pathogenesis

果蝇作为研究 Rett 发病机制的模型

• 批准号: 6800378
• 负责人: HOLLY N CUKIER
• 金额: $ 3.09万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6800378
• 关键词: CpG islands; Drosophilidae; Rett syndrome; arthropod genetics; biotechnology; disease /disorder model; gene expression; genetic screening; genetically modified animals; interdisciplinary collaboration; model design /development; molecular pathology; nucleic acid sequence; pathologic process; phenotype; predoctoral investigator

DESCRIPTION (provided by applicant): Mutations in the gene encoding methyl-CpG-binding protein 2 (MeCP2) cause Rett syndrome (RTT) and a host of other mental retardation syndromes in both males and females. RTT results from loss-of-function mutations of MECP2, which binds to target DNA and recruits Sin3A and histone deacetylases to silence transcription. MeCP2 is part of a larger family of genes, the methyl-CpG-binding domain (MBD) family, which is conserved from fly through humans. Overexpression of MECP2 in Drosophila using the UAS-GAL4 system produces a phenotype, possibly by competing with MBD genes. I propose to continue characterization of the full-length lines and create additional transgenic flies: two lines which overexpress MECP2 mutations commonly found in Rett patients and one line which produces MeCP2 without the MBD region. Secondly, I will characterize the Drosophila MBD gene CG10042 for native RNA and protein expression patterns. Then I will investigate its function through P-element mutagenesis and RNAi with genomic eDNA hybrids. I will characterize all resulting phenotypes and use these strains to identify modifiers of MeCP2 and/or CG10042 through an F 1 modifier screen to gain insight into pathways of MeCP2 function, which until now has been studied only in vitro. Through the proposed genetic studies, I hope to identify proteins that can be then studied to investigate the mechanisms of the mental and neurological dysfunction in Rett and related disorders.

描述（由申请人提供）：编码甲基 CpG 结合蛋白 2 (MeCP2) 的基因突变会导致男性和女性出现雷特综合征 (RTT) 和许多其他智力低下综合征。 RTT 是 MECP2 功能丧失突变的结果，MECP2 与靶 DNA 结合并招募 Sin3A 和组蛋白脱乙酰酶来沉默转录。 MeCP2 是甲基 CpG 结合域 (MBD) 家族这一更大基因家族的一部分，该家族从果蝇到人类都是保守的。使用 UAS-GAL4 系统在果蝇中过度表达 MECP2 可能会通过与 MBD 基因竞争来产生表型。我建议继续对全长品系进行表征并创建额外的转基因果蝇：两种品系过度表达常见于 Rett 患者中的 MECP2 突变，另一种品系产生不带 MBD 区域的 MeCP2。其次，我将表征果蝇 MBD 基因 CG10042 的天然 RNA 和蛋白质表达模式。然后我将通过 P 元件诱变和基因组 eDNA 杂交体的 RNAi 来研究其功能。我将表征所有产生的表型，并使用这些菌株通过 F 1 修饰物筛选来识别 MeCP2 和/或 CG10042 的修饰物，以深入了解 MeCP2 功能的途径，而迄今为止，该途径仅在体外进行了研究。通过拟议的基因研究，我希望鉴定出可用于研究雷特精神和神经功能障碍及相关疾病的机制的蛋白质。