CAREER: Optogel Proteomics: unbiased subcellular proteomics powered by photoreactions in hydrogel

职业：Optogel 蛋白质组学：由水凝胶中的光反应驱动的无偏亚细胞蛋白质组学

• 批准号: 2341058
• 负责人: Xiaoyu Shi
• 金额: $ 67.98万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-07-01 至 2029-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2341058&HistoricalAwards=false
• 关键词: CAREER; Optogel; Proteomics; unbiased; subcellular

Imagine cells as complex cities, where proteins are the citizens, each with specific roles in different districts. For scientists to understand how these cellular cities function, they need to map where each protein citizen lives and works. However, this is quite a challenge because current scientific tools are either not detailed enough or only focus on a few proteins at a time. This NSF CAREER project introduces a new method named Optogel Proteomics, which acts like a high-tech GPS to track proteins with remarkable precision that is smaller than a single cell. By using hydrogels and a powerful microscope, the investigators will isolate and study proteins from exact locations within the cell. The first mission is to explore the nuclear lamina, a meshwork wrapping a cell's nucleus that can change due to cancer or aging, to discover what proteins are present and how they may influence cancer progression and the aging process. This project not only aims to pioneer this new technology but also to share it widely, enhancing research in biology and engineering. Additionally, the project’s leader, a woman scientist, is dedicated to bridging the gender gap in science through special camps and workshops, empowering the next generation of female scientists. This NSF CAREER project seeks to overcome the limitations of subcellular spatial proteomics through the development of Optogel Proteomics. By employing expansion microscopy, photoreactions, and multi-photon microscopy, proteins within precise subcellular locations can be isolated and subsequently analyzed via mass spectrometry (MS). The project's objective is to provide an unbiased, high-resolution proteomic analysis of subcellular structures, with an initial focus on the nuclear lamina, which has been implicated in cancer-related and age-related heterochromatin loss. The novel Optogel Proteomics will be validated against the newly developed super-resolution microscopy method, Label-Retention Expansion Microscopy (LR-ExM). The broader impacts include the dissemination of Optogel Proteomics and LR-ExM, enhancing research capacity in life sciences at minimal costs. The principal investigator will also conduct annual workshops on these spatial analysis technologies and promote gender equity and interdisciplinary science exposure.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

想象细胞为复杂的城市，其中蛋白质是公民，每个城市都具有不同程度的特定作用。为了使科学家了解这些细胞城市的运作方式，他们需要绘制每个蛋白质公民的生活和工作的位置。但是，这是一个挑战，因为当前的科学工具要么不够详细，要么一次专注于几种蛋白质。这个NSF职业项目推出了一种名为Optogel蛋白质组学的新方法，该方法像高科技的GPS一样，可跟踪蛋白质，其精度高于单个单元格。通过使用水凝胶和强大的显微镜，研究人员将从细胞内的精确位置分离并研究蛋白质。第一个任务是探索核薄片，这是一种包裹细胞核的网状功能，该核能可能会因癌症或衰老而改变，以发现存在哪些蛋白质以及它们如何影响癌症进展和衰老过程。该项目不仅旨在开拓这项新技术，而且还旨在广泛分享它，从而增强生物学和工程学的研究。此外，该项目的领导者是一名女科学家，致力于通过特殊营地和讲习班来弥合科学中的性别差距，从而赋予下一代女性科学家的能力。这个NSF职业项目旨在通过开发Optogel蛋白质组学来克服亚细胞空间蛋白质组学的局限性。通过采用膨胀显微镜，光反应和多光子显微镜，可以分离精确的亚细胞位置内的蛋白质，然后通过质谱法（MS）分析并随后分析。该项目的目标是对亚细胞结构提供无偏的高分辨率蛋白质组学分析，最初的重点是核层，这与癌症相关且与年龄有关的异染色质损失有关。新型的Optogel蛋白质组学将通过新开发的超分辨率显微镜方法，标签 - 滞留显微镜（LR-EXM）验证。更广泛的影响包括传播Optogel蛋白质组学和LR-EXM，以最低的成本增强了生命科学的研究能力。首席研究者还将对这些空间分析技术进行年度研讨会，并促进性别平等和跨学科科学曝光。该奖项反映了NSF的法定任务，并通过使用基金会的智力优点和更广泛的影响来评估NSF的法定任务。