Synthetic Biology and Optogenetics Core

摘要 该计划项目将在所有项目中使用新型工程试剂，使操作和 通过精确的空间和时间控制对单个信号通路进行询问。如个别内容所述 项目，核心B提供的试剂将用于定义调节可塑性和 巨噬细胞的特化，并确定它们在肺部炎症和损伤中的作用以及解决。 这些工具的开发和优化将需要合成生物学和光遗传学的努力 核心B的主要功能是：1)为每个特定的特定目标开发定制的分子工具 个别项目中的问题，2)在新试剂上线时对其进行评估，并为 它们在巨噬细胞和活体小鼠模型中的应用，以及3)为所有项目提供援助 工具的应用、故障排除和结果分析。其目的是简化应用程序 在这些先进的技术中，剖析计划项目中的特定信号通路并允许 与会者集中讨论需要这些试剂的拟议问题。将开发的特定工具 核心B使用的包括：1)新试剂来控制识别的蛋白质的定位和相互作用 通过项目；以及2)调节活巨噬细胞中选定蛋白质活性的试剂和靶向 这些蛋白质在特定的复合体和亚细胞位置被激活。核心B还将生成和 优化巨噬细胞表达工程蛋白的试剂，并根据需要开发新的试剂 用于操纵和询问蛋白质相互作用和细胞信号。开发的工具将使 单个信号通路的操纵、这些通路的解构以及它们的作用的评估 肺内巨噬细胞的调节功能。

Abstract The Program Project will employ in all projects novel engineered reagents that enable manipulation and interrogation of individual signaling pathways with precise spatial and temporal control. As described in individual Projects, reagents provided by Core B will be used to define the signaling processes that regulate plasticity and specialization of macrophages and define their role during lung inflammation and injury as well as resolution. Development and optimization of these tools will require the efforts of the Synthetic Biology and Optogenetics Core B. The central functions of Core B will be 1) to develop molecular tools customized for each specific question in individual Projects, 2) to evaluate the new reagents as they come on-line and establish protocols for their application in macrophages and in in vivo mouse model, and 3) to provide assistance to all Projects with the application of the tools, troubleshooting, and analysis of the results. The intent will be to simplify application of these advances technologies to dissect specific signaling pathways in the Program Project and allow participants to focus on the proposed questions requiring these reagents. The specific tools that will be developed and employed by Core B include: 1) new reagents to control localization and interactions of proteins identified by Projects; and 2) reagents for regulation of activity of selected proteins in living macrophages, and targeted activation of these proteins in specific complexes and subcellular locations. Core B will also generate and optimize reagents for expression of engineered proteins in macrophages and develop new reagents as needed for manipulation and interrogation of protein interactions and cell signaling. The tools developed will enable manipulation of individual signaling pathways, deconstruction of these pathways, and assessment of their roles in the regulating function of macrophages in lungs.