Bioorthogonal temporospatial tools

项目摘要 生命系统通过精确的控制--地点、时间、动力学和强度--编排分子事件 - 并在突触网络、基因表达、表观遗传中记录这些事件的记忆。 标记，以及无数其他控制生物反应的时间和地点的电路。想象一下 编排和追踪这些历史是化学家和生物学家可以用 只有部分准确性，相当大的误差，和有限的时间范围。因此，我们的研究议程 专注于构建新的化学工具，用于生命系统的时空分析， 围绕部署下一级生物正交所产生的紧急属性进行组织 多层（生物）分子结构内的化学。由此产生的混合动力系统规避 长期的挑战，实现：i）同时的速度/稳定性，高效的实时分子 ii）检测罕见/独特事件的灵敏度;以及 iii）特异性/多样性，用于准确检测和细粒度分子编码（亚）细胞 穿越时间的历史 基于正在进行的机理研究的势头和我们最近的实验成功， 为了实现活细胞和组织的多路成像，我们在未来五年的目标是： 在利用二维/三维拓扑结构的应用中的生物正交化学，而不是 单个连接/切割事件，以及在利用核酸杂交以 编码序列识别，加速反应动力学，并使信号放大。一个集合中 项目，我们的目标是创建自放大可编程生物正交反应，阐述 这个新工具包的能力，并应用它们来改变我们的方法，可视化活细胞， 组织中在另一个实验中，我们设想了序列生成架构， 将化学反应转化为可放大的生物信息，建立生物正交的概念 翻译和螯合性。与容易融入现有高- 性能核酸生物技术，我们预计广泛的适用性和快速下游 开发新一代工具，用于跟踪（生物）分子、单个细胞和群体。

PROJECT SUMMARY Living systems choreograph molecular events with precise control—place, time, kinetics, and intensity —and record memories of those occurrences in synaptic networks, gene expression, epigenetic marks, and myriad other circuits that govern the where&when of biologic responses. Visualizing this choreography and tracing these histories are tasks that chemists and biologists can accomplish with only partial accuracy, considerable effort, and limited temporal range. Our research agendas are thus focused on constructing new chemical tools for temporospatial analysis of living systems, and organized around the emergent properties that result from deploying next-level bioorthogonal chemistries within multi-layered (bio)molecular architectures. The resulting hybrid systems circumvent perennial challenges, achieving: i) simultaneous speed/stability, for efficient real-time molecular machinery and longitudinal performance in vivo; ii) sensitivity for detection of rare/unique events; and iii) specificity/multiplicity, for accurate detection and fine-grained molecular encoding of (sub)cellular histories across time. Building on the momentum of ongoing mechanistic investigations and the success of our recent effort to achieve multiplexed imaging of living cells and tissues, our goals for the next five years extend bioorthogonal chemistry in applications that exploit two/three dimensional topologies, rather than singular ligation/cleavage events, and in architectures that leverage nucleic acid hybridization to encode sequence recognition, accelerate reaction kinetics, and enable signal amplification. In one set of projects, we aim to create self-amplifying programmable bioorthogonal reactions, elaborate the capabilities of this new toolkit, and apply them to transform our methods for visualizing living cells and tissues. In another, we have envisioned sequence-generating architectures that convert biocompatible chemical reactivity into amplifiable biological information, establishing the concepts of bioorthogonal translation and sequegenicity. With scaffolds that readily integrate into the workflows of existing high- performance nucleic acid biotechnologies, we anticipate broad applicability and rapid downstream development of a new generation of tools for tracking (bio)molecules, individual cells, and populations.