Bioorthogonal temporospatial tools

生物正交时空工具

• 批准号: 10711005
• 负责人: Jonathan Carlson
• 金额: $ 41.75万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711005
• 关键词: 3-Dimensional; Acceleration; Architecture; Biological; Biotechnology; Cells; Chemicals; Chemistry; Detection; Development; Epigenetic Process; Event; Gene Expression; Generations; Goals; Grain; Hybrids; Individual; Investigation; Kinetics; Ligation; Memory; Methods; Molecular; Nucleic Acid Hybridization; Nucleic Acids; Organism; Performance; Population; Property; Reaction; Recording of previous events; Research; Signal Transduction; Specificity; Speed; Synapses; System; Time; Tissues; Translating; Translations; Visualization; biomaterial compatibility; detection sensitivity; in vivo; molecular dynamics; multiplexed imaging; response; scaffold; success; tool

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.

项目摘要 生活系统编舞分子事件具有精确控制 - 位置，时间，动力学和强度 - 并记录突触网络中这些发生的记忆，基因表达，表观遗传 标记和其他众多的圈子，这些圈子控制着生物学反应的位置和时间。可视化这个 编排和追踪这些历史是化学家和生物学家可以完成的任务 仅部分准确性，相当大的效果和有限的临时范围。因此，我们的研究议程是 专注于构建新的化学工具，用于暂时的生活系统，以及 由部署下一级生物串管造成的新兴特性组织 多层（BIO）分子体系结构中的化学成分。由此产生的混合系统绕过 多年生挑战，实现：i）简单的速度/稳定性，用于有效的实时分子 体内机械和纵向性能； ii）检测稀有/独特事件的灵敏度；和 iii）特殊的城市/多样性，以实现（子）细胞的精确检测和细粒度的分子编码 跨时间的历史。 以持续的机械调查和我们最近的作用的成功为基础 为了实现活细胞和组织的多路复用成像，我们接下来的五年的目标延长了 在利用两/三维拓扑的应用中的生物正交化学，而不是 奇异的连接/切割事件，以及在利用核酸杂交的体系结构中 编码序列识别，加速反应动力学和启用信号扩增。一组 在项目中，我们旨在创建自我扩增的可编程生物反应，并详细阐述 这个新工具包的功能，并将其应用于我们的方法，以可视化活细胞和 组织。在另一个中，我们设想了序列生成的结构，这些结构转化了生物相容性 化学反应性对可扩增的生物学信息，建立生物正交的概念 翻译和sequegenicity。随着sca效应，很容易整合到现有高级的工作流中 性能核酸生物技术，我们预计适用于广泛的适用性和快速的下游 开发新一代的跟踪工具（BIO）分子，单个细胞和人群。