Uncovering new approaches to manipulate and probe biochemical processes with spatial and temporal precision.

发现以空间和时间精度操纵和探测生化过程的新方法。

• 批准号: 10581649
• 负责人: Chandra L Tucker
• 金额: $ 43.04万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581649
• 关键词: Area; Biochemical Process; Biological; Biological Process; Cell physiology; Cells; Disease; Engineering; Environment; Genetic; Goals; Health; Imaging technology; Light; Outcome; Photoreceptors; Plant Photoreceptors; Protein Dynamics; Protein Engineering; Proteins; Regulation; Research; Scientist; Testing; Time; Work; cryptochrome 2; engineering design; improved; novel; novel strategies; optogenetics; protein complex; protein function; tool

While advances in imaging technologies are enabling scientists to observe cellular phenomena and protein complexes in increasingly greater detail, our ability to perturb and manipulate cells with equally fine precision have lagged. To better understand the dynamic cellular environment, tools that can selectively perturb biological processes, in real-time, with fast, reversible, tunable, and spatially- selective control are required. A major step forward in this area has come from the field of cellular optogenetics, wherein photosensory proteins are engineered to control various cellular processes with light. Because light can be delivered with fast temporal and subcellular spatial precision, such tools allow unprecedented biological control. In prior work, we developed novel optogenetic tools based on the plant photoreceptor cryptochrome 2 (CRY2), which can be used to precisely manipulate protein interactions, localization, and activity. While these tools have been successful at regulating protein function, improvements and new strategies for optogenetic regulation are needed. In the proposed work, we will usher in a new wave of optogenetic engineering, improving upon existing strategies, extending optogenetic tools into new areas, developing new photoreceptor modules, and testing novel engineering designs to regulate protein function with light. The long term goals of the project are to enable a set of modular precision tools for probing and manipulating biochemical processes with fast temporal control and over spatial scales ranging from subcellular to organismal.

虽然成像技术的进步使科学家能够观察细胞现象， 蛋白质复合物越来越多的细节，我们的能力，扰乱和操纵细胞与 同样精细的精度也落后了。为了更好地了解动态细胞环境， 可以选择性地干扰生物过程，实时，快速，可逆，可调，空间- 需要选择性控制。这一领域的一个重大进展来自蜂窝领域， 光遗传学，其中光敏蛋白被工程化以控制各种细胞过程， 光因为光可以以快速的时间和亚细胞空间精度被递送，所以这样的工具 前所未有的生物控制。在先前的工作中，我们开发了基于 植物光感受器隐花色素2（Photoreceptor cryptochrome 2，简称CP 2），可用于精确操纵蛋白质 互动、定位和活动。虽然这些工具已经成功地调节蛋白质 需要光遗传学调节的功能、改进和新策略。拟议 工作，我们将迎来新一波的光遗传工程，改善现有的战略， 将光遗传学工具扩展到新的领域，开发新的光感受器模块，并测试新的 工程设计来调节蛋白质的功能与光。该项目的长期目标是 使一套模块化的精密工具，用于探测和操纵生化过程， 时间控制和空间尺度，从亚细胞到有机体。