Mechanisms regulating tau alternative pre-mRNA splicing

tau 选择性前 mRNA 剪接的调节机制

基本信息

批准号：
8311629
负责人：
JANE Y WU
金额：
$ 31.26万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-15 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8311629
关键词：
Address Affect Alternative Splicing Biochemical Bioinformatics Biological Biological Assay Cell Culture System Cell Death Cells Complex DNA Sequence Defect Dementia Disease Dissection Elements Enhancers Exons Frontotemporal Dementia Gene Mutation Genes Genetic Goals Human In Vitro Lead Mass Spectrum Analysis Methods Molecular Molecular Genetics Mutation Nerve Degeneration Neurodegenerative Disorders Neuronal Differentiation Neurons Pathogenesis Patients Pattern Play Proteins Proteomics Publishing RNA Interference RNA Processing RNA Splicing Reagent Regulation Regulator Genes Reporting Research Risk Factors Role Sampling Screening procedure Spliced Genes Structure Surveys System Tauopathies Temporal Lobe Testing Tissue Sample Trans-Activators Transcript Work base clinical phenotype crosslink expression cloning gene function mRNA Precursor nervous system disorder neuron loss neuronal survival stem tau Proteins tau expression tau mutation tool

项目摘要

DESCRIPTION (provided by applicant): Mechanisms regulating tau alternative pre-mRNA splicing Accumulating evidence indicates that pre-mRNA splicing defects and aberrant splicing lead to a range of neurodegenerative disorders. Alternative splicing is critical for tau gene function, and splicing mutations in the human tau gene lead to fronto-temporal lobe dementia (FTLD-tau). Although tau splicing mutations were initially reported in 1998, the mechanisms by which mutations affect tau alternative splicing are still poorly understood, and risk factors as well as genetic modifiers in FTLD-tau remain to be identified. This proposal aims to investigate the mechanisms regulating tau gene alternative splicing. We plan to use molecular, biochemical and cell biological methods to investigate tau alternative splicing and to compare the differential spliceosomal recognition of wild type versus mutant tau transcripts. We have established both in vitro biochemical assays and cell culture systems to investigate tau exon 10 alternative splicing. Using biochemical assays and an expression cloning strategy, we have identified several proteins involved in tau exon 10 alternative splicing in our preliminary studies. A primary neuronal culture system has also been set up to investigate the role of identified factors in neuronal cell death. Molecular dissection of cis- and trans-acting elements important for tau alternative splicing will not only help in understanding the basic mechanisms controlling tau alternative splicing but also reveal new players in pathogenesis of neurodegeneration. We plan to use combined bioinformatics, biochemical, molecular and genetic approaches to study the pathogenetic mechanisms underlying FTLD-tau, one most common neurodegenerative disorders.

描述（由申请人提供）：调节TAU替代性mRNA剪接的机制积累证据表明，前mRNA剪接缺陷和异常剪接导致一系列神经退行性疾病。替代剪接对于tau基因功能至关重要，人tau基因的剪接突变导致额叶叶型痴呆（FTLD-TAU）。尽管最初在1998年报道了tau剪接突变，但突变影响tau替代剪接的机制仍然很少了解，危险因素以及FTLD-TAU中的遗传修饰符仍有待鉴定。该建议旨在研究调节tau基因替代剪接的机制。我们计划使用分子，生化和细胞生物学方法来研究TAU替代剪接，并比较野生型与突变体Tau转录本的差异剪接识别。我们已经建立了体外生化测定和细胞培养系统，以研究TAU外显子10替代剪接。使用生化测定和表达克隆策略，我们在初步研究中确定了几种参与TAU外显子10替代剪接的蛋白质。还建立了原发性神经元培养系统，以研究已鉴定因子在神经元细胞死亡中的作用。顺式和反式作用元件的分子解剖对TAU替代剪接重要的元素不仅有助于理解控制TAU替代剪接的基本机制，而且还揭示了神经变性的发病机理的新参与者。我们计划使用合并的生物信息学，生化，分子和遗传方法来研究FTLD-TAU的致病机制，这是最常见的神经退行性疾病。