RIG: The Role of POT1 in Telomere Maintenance and Protection

RIG：POT1 在端粒维持和保护中的作用

• 批准号: 0721463
• 负责人: Diego Loayza
• 金额: $ 15万
• 依托单位: CUNY Hunter College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-15 至 2011-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0721463&HistoricalAwards=false
• 关键词: RIG; Role; POT1; Telomere; Maintenance

Telomeres are essential for chromosome stability because they provide protection to chromosome ends, which would otherwise be detected as DNA breaks, and they ensure their proper replication. The main hypothesis of this project is that POT1, the overhang binding protein at telomeres, regulates both telomerase-mediated telomere maintenance and telomere protection. A molecular genetic analysis of POT1 function will be carried out to elucidate how this regulation is achieved. To that end, the investigator will construct a conditional POT1 knockout in somatic cells, thereby providing a clean genetic system in which to study POT1 function. The strategy entails introducing LoxP recombination sites on either side of a POT1 exon in order to induce the POT1-null phenotype at will upon expression of Cre recombinase. This work will produce an experimental system tailored for the study of the short-term and long-term effects of POT1 loss, and for detecting any rescue of the cells by specific alleles or domains of POT1 and other candidate proteins. The project is well-suited to the training of students at the beginning of theirscientific careers. It will teach them the subtleties of molecular genetic analysisand the possible impact of specific pathways on cell viability. The student body at Hunter College is one of the most diverse in the nation, and is largely undergraduate. Undergraduates will have the opportunity to be directly involved in modern molecular biological research, while taking classes in cell biology, biochemistry and genetics.

端粒对染色体稳定性至关重要，因为它们为染色体末端提供保护，否则会被检测为DNA断裂，并且它们确保其正确复制。该项目的主要假设是POT1，端粒的突出端结合蛋白，调节端粒酶介导的端粒维持和端粒保护。将进行POT1功能的分子遗传学分析，以阐明这种调节是如何实现的。为此，研究人员将在体细胞中构建条件性POT1敲除，从而提供一个干净的遗传系统来研究POT1功能。该策略需要在POT1外显子的任一侧引入LoxP重组位点，以便在Cre重组酶表达时随意诱导POT1无效表型。这项工作将产生一个专门用于研究POT1丢失的短期和长期影响的实验系统，并用于检测POT1和其他候选蛋白的特定等位基因或结构域对细胞的任何拯救。该项目非常适合于对刚开始科学生涯的学生进行培训。它将教会他们分子遗传分析的微妙之处以及特定途径对细胞活力的可能影响。亨特学院的学生是全国最多样化的学生之一，大部分是本科生。 本科生将有机会直接参与现代分子生物学研究，同时参加细胞生物学，生物化学和遗传学课程。