Collaborative Research: Harnessing plant hormone receptors for the rapid design of genetic circuits controlled by user-specified ligands

生命之树上的有机体可以感知其环境的特征并对其做出反应。例如，植物向着光的方向生长，细菌朝着营养来源游去。生物体的感知和反应能力是一种核心的生物功能，由分子机制支持，它识别特定的信号，然后触发反应，如生物体的生长、发育或运动的变化。在这个项目中，一组工程师和生物学家一起工作，通过设计一个从植物中提取的感觉-反应模块来开发新的生物功能。这种传感器是独一无二的，因为它可以用作一个简单的开关，随意打开和关闭其他蛋白质，并可以重新编程，以识别不同的化学物质。该项目开发了用于药物和饮食分子的传感器，然后使用这些新的传感器来设计可以很容易地指示许多不同分子的存在的合成“哨兵细胞”。例如，该项目设计的细胞会根据接触的药物而变成不同的颜色。为了做到这一点，该团队正在对植物传感器进行重新编程，以识别新分子，然后将新传感器与调节基因表达的细菌酶物理联系起来。这种结合使细菌能够根据输入信号改变不同的颜色。该项目支持学生在STEM职业生涯中的培训，研究成果被整合到一个以发现为基础的实验室中，并与一门入门实验室课程相关联。该项目能够快速构建由用户指定的分子控制的单通道和多通道遗传电路。这种能力是通过建立一个快速构建可编程化学诱导的RNA聚合酶(Chirp)的平台来实现的。为此，使用计算设计、突变和遗传选择来重新编程一种新的植物衍生的化学诱导二聚模块的配体结合特异性，该模块用于调节分裂的T7 RNA聚合酶的活性。同时，一组识别新启动子的新的T7正交变异体被开发并转化为啁啾，以提供多输入/多输出的化学调节电路。这些努力开启了一系列新的配体控制的生物技术，并允许研究人员为目标应用设计由最佳特定配体控制的复杂遗传电路。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Organisms across the tree of life can sense features of their environments and respond to them. For example, plants grow in the direction of light, and bacteria swim toward nutrient sources. An organism’s ability to sense and respond is a core biological function undergirded by molecular machinery that recognizes specific signals and then triggers responses, such as changes in the organism’s growth, development, or movement. In this project, a team of engineers and biologists work together to develop new biological functions by engineering a sense-response module taken from plants. This sensor is unique because it can be used as a simple switch for turning other proteins on and off at will and can be reprogrammed to recognize diverse chemicals. This project develops sensors for pharmaceuticals and dietary molecules and then uses these new sensors to design synthetic ‘sentinel cells’ that can easily indicate the presence of many different molecules. For example, the project designs cells that turn different colors depending on to which pharmaceutical they are exposed. To do this, the team is reprograming a plant sensor to recognize new molecules and then physically link the new sensors to a bacterial enzyme that regulates gene expression. This union enables bacteria to turn different colors in response to input signals. The project supports the training of students in STEM careers and the research findings are integrated into a discovery-based lab associated with an introductory laboratory course.This project enables the rapid construction of single and multi-channel genetic circuits controlled by user-specified molecules. This capacity is enabled by building a platform for the rapid construction of programmable chemically inducible RNA polymerases (ChIRPs). To do this, computational design, mutagenesis, and genetic selections are used to reprogram the ligand-binding specificity of a novel plant-derived chemical-induced dimerization module that is used to regulate the activity of split T7 RNA polymerase. In parallel, a new set of orthogonal T7 variants that recognize novel promoters are developed and converted into ChIRPs to deliver multi-input/multi-output chemical-regulated circuitry. These efforts open a vast range of new ligand-controlled biotechnologies and allow investigators to design complex genetic circuits controlled by the best specific ligands for target applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.