From synthetic bacterial adhesions to synthetic bacterial materials

细菌合成多细胞系统和材料的工程为许多健康带来了希望- 相关应用，如模块化药物生物合成、活诊断设备和合成生物film研究。 模特们。到目前为止，细菌合成生物学主要集中在分子和单细胞的规模上。 关于细菌合成联合体的同等工作要落后得多，这在很大程度上是因为之前缺乏fi 合适的合成和基因编码的细胞-细胞黏附工具来控制组装、发育和 多细胞系统的功能。我们最近开发了fi第一个合成细胞-细胞黏附工具箱，例如 以及用于光遗传控制细胞表面沉积和图案化的工具。 这项研究的特殊目标是显著地促进这些合成细胞黏附工具的发展，以及 开发设计原则和预测性建模工具，使联盟能够进行工程设计和构图 整合所有相关的长度尺度(即分子、细胞和多细胞)，并最终为 与医学相关的应用。我们的主要假设是，我们可以显著地提高对fi的控制 在大肠杆菌中合成黏附蛋白的强度、特殊fi城市和亚细胞定位，这将 允许合理调整联合体级别的生物物理属性，如孔隙度和粘弹性，并且 最终将实现多功能多细胞联合体工程和图案化。这项工作将构成 各种生物医学应用的基础，如生物兼容材料、多细胞即插即用 途径工程、靶向体内药物输送和活的诊断设备。 我们的跨学科方法结合了合成生物学、生物物理学、仪器仪表和建模。全 实验将以定量的方式进行。拟议的调查包括三项独立的调查 由我们的假设激发的协同fic目标：(目标1)促进合成粘附素的功能 亚细胞级别的工具包；(目标2)实现对合成联合体属性的工程控制，如 粘弹性和孔隙率在10-100微米范围内；以及(目标3)实现更高级别的联合体图案 在厘米的尺度上，并展示了医疗应用的潜力。 PI(里德尔-克鲁斯教授)和他的团队非常适合这个项目，因为我们有重要的fiCan 在合成生物学、生物物理学、仪器设备(例如，微fl指南、成像)和基因建模方面的专业知识 不同尺度的电路和生物物理系统。我们开发了fi第一个合成细胞-细胞和光遗传细胞表面 细菌中的黏附工具箱。多个协作者在关键领域提供额外的领域专业知识。总的来说， 该项目的创新之处在于将合成粘附素建立为 综合电路工程工具箱和建立多细胞模块化工程的新范例 活体材料。因此，该项目将对合成财团的工程产生广泛影响 研究以及实现未来健康应用的动态频谱。

Engineering of bacterial synthetic multicellular systems and materials hold promise for many health- relevant applications such as modular drug biosynthesis, living diagnostic devices, and synthetic biofilm research models. To date, bacterial synthetic biology has largely focused on the scales of molecules and single cells. Equivalent work on bacterial synthetic consortia is much less advanced, in significant part due to the previous lack of suitable synthetic and genetically encoded cell-cell adhesion tools to control the assembly, development, and functionality of multicellular systems. We recently developed the first such synthetic cell-cell adhesion toolbox, as well as tools for optogenetically controlling cell-surface deposition and patterning. The specific objectives of this research are to significantly advance these synthetic cell-adhesion tools, and to develop design principles and predictive modeling tools that enable consortia engineering and patterning that integrate all relevant length scales (i.e., molecular, cellular, and multicellular), and ultimately pave the way for medially relevant applications. Our main hypothesis is that we can significantly advance our control over the strength, specificity, and subcellular localization of synthetic adhesion proteins in Escherichia coli, which will allow rational tuning of consortium-level biophysical properties such as porosity and viscoelasticity, and which will ultimately enable versatile multicellular consortium engineering and patterning. This work will constitute a foundation for various biomedical applications such as biocompatible materials, multicellular plug-and-play pathway engineering, targeted in-vivo drug delivery, and living diagnostic devices. Our interdisciplinary methodology combines synthetic biology, biophysics, instrumentation and modeling. All experiments will be done in a quantitative manner. The proposed investigations include three independent yet synergistic Specific Aims motivated by our hypothesis: (Aim 1) Advance the functionality of the synthetic adhesin toolkit at the subcellular level; (Aim 2) Achieve engineering control over synthetic consortium properties such as viscoelasticity and porosity at the scale of 10-100 µm; and (Aim 3) Achieve higher-level consortium patterning on the scale of centimeters and demonstrate potential for medical applications. The PI (Prof. Riedel-Kruse) and his team are well-suited for this project as we have significant expertise in synthetic biology, biophysics, instrumentation (e.g., microfluidics, imaging), and modeling genetic circuits and biophysical systems across scales. We developed the first synthetic cell-cell and optogenetic cell-surface adhesion toolboxes in bacteria. Multiple collaborators provide additional domain expertise in key areas. Overall, this project's innovation lies in establishing synthetic adhesins as an essential and integral component of the synthetic circuit-engineering toolbox and in establishing a novel paradigm for modular engineering of multicellular living materials. Accordingly, this project will broadly impact the engineering of synthetic consortia for basic research as well as enable a dynamic spectrum of future applications in health.