A multifunctional mitochondrial genetic probe to understand mitochondrial biology.

用于了解线粒体生物学的多功能线粒体遗传探针。

• 批准号: 2106095
• 金额: --
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2106095
• 关键词: multifunctional; mitochondrial; genetic; probe; understand

The Stagi lab has developed a set of transfectable/transducable probes which allow the study of mitochondria in biochemical, proteomic and physiological applications. Briefly, the probe is genetically encoded and incorporated in the mitochondria. The set is composed of: Mitochondrial markers in different colours recruited to outer or inner membranes with localized biotinylation systems for spin-free isolation. Coupled to this are probes with Ca2+, pH and membrane potential sensors which allow live imaging, and FACS sorting so that mitochondria with specific characteristics can be isolated. This allows coupled live imaging and proteomic applications in cell lines. The project will involve optimising techniques to isolate and study mitochondria, using the state of the art of imaging, biochemistry enzymology and physiology, to develop robust assays for specific mito-proteomics/metabolomics studies. We will compile proteomic assays of mitochondria isolated with this new innovative biochemical system. If the testing and completion of this set of tools will be successful is certainly have a large application in study the physiology of mitochondria directly in the organelle without contamination.

Stagi实验室开发了一套可转染/可转导的探针，可以在生化，蛋白质组学和生理学应用中研究线粒体。简言之，探针被遗传编码并掺入线粒体中。该套件包括：不同颜色的线粒体标记物通过局部生物素化系统被募集到外膜或内膜，以进行无自旋分离。与此耦合的是具有Ca 2+、pH和膜电位传感器的探针，其允许活体成像和FACS分选，使得可以分离具有特定特征的线粒体。这允许在细胞系中耦合活体成像和蛋白质组学应用。该项目将涉及优化分离和研究线粒体的技术，利用成像，生物化学酶学和生理学的最新技术，为特定的线粒体蛋白质组学/代谢组学研究开发强大的检测方法。我们将编译用这种新的创新生化系统分离的线粒体的蛋白质组学测定。如果这套工具的测试和完成将是成功的，肯定是有很大的应用在研究线粒体的生理直接在细胞器中没有污染。