Light-Actuatable NanoRobots for Molecular Uncaging

描述(由申请人提供) 摘要：精确的时空控制配体进入完整的细胞、器官和生物体，对于神经递质、神经调节剂和激素在复杂生物系统(如大脑)的运行中的功能的时间分辨和因果分析是重要的。在过去的几十年里，通过结合亲光化学基团来封闭几个小分子已经取得了一些成功，这些基团在没有光的情况下阻断了分子的生物活性，并在紫外光照射下释放出生物活性分子。然而，这种光化学修饰对于小分子来说是困难的，对于活性中心太大而不能被添加的化学基团阻止的大分子来说几乎是不可能的。此外，笼子分子经常在发光之前就表现出泄漏的生物活性，而紫外线的使用可能会破坏细胞成分；因此，尽管在体外证明了有效性，但在体内很少使用去膜来研究完整的生物系统。在这里，我将描述一种新的战略 能够以毫秒级的时间分辨率去除任意的生物活性分子和多肽，使用可见光在体内使用是安全的。我将使用计算蛋白质和DNA设计来构建光可控的纳米机器人，并使用这些纳米机器人在完整的器官系统中连接各种分子、多肽和蛋白质。这种策略有可能以前所未有的时空精度彻底改变配体功能的研究，在基础分子和系统神经科学、药物开发和副作用评估方面开辟新的前沿。

DESCRIPTION (Provided by the applicant) Abstract: Precise spatiotemporal control of ligand delivery into intact cells, organs, and organisms is important for the time resolved and causal analysis of the functions of neurotransmitters, neuromodulators, and hormones in the operation of complex biological systems, such as the brain. Over the past several decades, some success has been achieved in uncaging several small molecules through the attachment of photolabile chemical groups, which block the bioactivity of the molecule in the absence of light and release a bioactive molecule upon UV light irradiation. However, such photochemical uncaging modifications are difficult to develop for small molecules, and are nearly impossible for large molecules whose active sites are often too large to be blocked by the addition of chemical groups. In addition, caged molecules often exert leak bioactivity even before light delivery, and the use of UV light can be damaging to cellular components; as a result, despite proven utility in vitro, uncaging has been little used in vivo to study intact biological systems. Here, I will describe a novel strategy capable of uncaging arbitrary bioactive molecules and peptides with millisecond time scale resolution, using visible light safe for in vivo use. I will use computational protein and DNA design to construct light controllable 'NanoRobots', and use these NanoRobots to uncage a variety of molecules, peptides, and proteins in the intact organ systems. Such a strategy has the potential to revolutionize the study of ligand functions with unprecedented spatiotemporal precision, opening up new frontiers in basic molecular and systems neuroscience, pharmaceutical development, and side effect assessment.