Germline Stem Cell Analysis Tools

生殖干细胞分析工具

• 批准号: 8947912
• 负责人: E. Jane Albert Hubbard
• 金额: $ 8.48万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-10 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8947912
• 关键词: Address; Animals; Biological Models; Caenorhabditis elegans; Cells; Color; Communication; Cues; Data; Degenerative Disorder; Distal; Educational process of instructing; Funding; Future; Genes; Genetic; Genetic Recombination; Germ Cells; Germ Lines; Goals; Human; Imagery; Individual; Investments; Label; Life; Ligands; Malignant Neoplasms; Mammals; Mediating; Modeling; Molecular; Monitor; Organism; Pathway interactions; Physiological; Physiology; Population; Population Dynamics; Regulation; Seeds; Signal Transduction; Stem cells; System; Techniques; Technology; Testing; Time; Transgenes; Transgenic Organisms; Undifferentiated; Ursidae Family; Whole Organism; Work; genetic manipulation; germline stem cells; interest; notch protein; public health relevance; recombinase; response; stem; stem cell biology; stem cell division; stem cell niche; stem cell technology; success; technology development; tool; transgene expression

 DESCRIPTION (provided by applicant): This project will develop, optimize and implement technology to lineally mark and/or genetically alter individual germ cells in C. elegans. The germ line in this worm shares many features in common with stem cell systems in all animals, including humans. For example, it is maintained by virtue of interaction with a cellular microenvironment or niche that produces ligands to activate a highly conserved Notch receptor on the germ cells. It was the first stem cell system for which a niche was defined at the cell and molecular levels. This stem cell system has since emerged as a powerful system to understand the dynamics of stem cell expansion, quiescence and plasticity in response to genetic, environmental and physiological alterations. The worm has a short (3-day) lifecycle, is transparent, short-lived, and highly amenable to environmental and genetic manipulation. Despite these advantages, understanding critical features of the germline stem cell system and its responses to environmental and genetic perturbations is currently limited by our relatively poor understanding of cellular mechanisms of renewal and how they are altered under experimental manipulation. To bridge this gap, we need to follow individual stem cells and their cellular progeny over time. The technology proposed here will bridge this gap. Taking advantage of recent progress in germline transgene expression in C. elegans, existing technologies from other organisms will be brought to the worm. In addition to lineage marking, the technology will make possible the manipulation of gene activity in single cells or subsets of cells. Uniquely for this model system, the technology will enable monitoring of germline stem cells in individuals over their entire lifetime and in conditions of whole-animal physiological perturbations. This advance will benefit the greater stem cell biology and germline fields as additional questions will become accessible in this premier model system.

 描述（由申请人提供）：该项目将开发、优化和实施线性标记和/或遗传改变秀丽隐杆线虫个体生殖细胞的技术。这种蠕虫的生殖系与包括人类在内的所有动物的干细胞系统具有许多共同特征。例如，它是通过与细胞微环境或生态位的相互作用来维持的，细胞微环境或生态位产生配体以激活生殖细胞上高度保守的Notch受体。这是第一个在细胞和分子水平上定义了利基的干细胞系统。此后，该干细胞系统已成为一个强大的系统，可以了解干细胞响应遗传、环境和生理变化的扩张、静止和可塑性的动态。该蠕虫的生命周期短（3 天），透明且寿命短，并且非常容易受到环境和基因操纵的影响。尽管有这些优点，但由于我们对细胞更新机制及其在实验操作下如何改变的了解相对较少，目前了解生殖干细胞系统的关键特征及其对环境和遗传扰动的反应受到限制。为了弥补这一差距，我们需要随着时间的推移跟踪单个干细胞及其细胞后代。这里提出的技术将弥补这一差距。利用线虫种系转基因表达的最新进展，其他生物体的现有技术将被引入线虫中。除了谱系标记之外，该技术还将使得操纵单个细胞或细胞亚群中的基因活性成为可能。该模型系统的独特之处在于，该技术将能够在个体的整个生命周期和整个动物的生理扰动条件下监测生殖系干细胞。这一进展将有利于更大的干细胞生物学和生殖系领域，因为在这个首要的模型系统中将可以解决更多问题。