From synthetic bacterial adhesions to synthetic bacterial materials

细菌合成多细胞系统和材料的工程化为许多健康- 相关应用，如模块化药物生物合成、活体诊断设备和合成生物膜研究 模型迄今为止，细菌合成生物学主要集中在分子和单细胞的尺度上。 关于细菌合成财团的同等工作进展要慢得多，这在很大程度上是由于之前的缺乏 合适的合成和遗传编码的细胞-细胞粘附工具，以控制组装，发展， 多细胞系统的功能。我们最近开发了第一个这样的合成细胞粘附工具箱， 以及用于光遗传学控制细胞表面沉积和图案化的工具。 这项研究的具体目标是显着推进这些合成细胞粘附工具， 开发设计原则和预测建模工具，使联盟工程和模式， 整合所有相关的长度尺度（即，分子、细胞和多细胞），并最终为 医学相关应用。我们的主要假设是，我们可以大大提高我们对 大肠杆菌中合成粘附蛋白的强度、特异性和亚细胞定位， 允许合理调整联合体水平的生物物理性质，例如孔隙度和粘弹性，并且 将最终实现多功能的多细胞联合体工程和模式化。这项工作将构成 生物相容性材料、多细胞即插即用等各种生物医学应用的基础 通路工程、靶向体内药物递送和活体诊断装置。 我们的跨学科方法结合了合成生物学，生物物理学，仪器和建模。所有 实验将以定量的方式进行。拟议的调查包括三个独立的，但 我们的假设激发的特定目标：（目标1）提高合成粘附素的功能 （目标2）实现对合成聚生体性质的工程控制， 粘弹性和孔隙率在10-100微米的规模;和（目标3）实现更高水平的财团图案化 以厘米为单位，并展示了医疗应用的潜力。 PI（Riedel-Kruse教授）和他的团队非常适合这个项目，因为我们有重要的 在合成生物学，生物物理学，仪器（例如，微射流、成像）和遗传建模 电路和生物物理系统。我们开发了第一个合成细胞-细胞和光遗传细胞-表面 细菌中的粘附工具箱。多个合作者在关键领域提供额外的领域专业知识。总的来说， 该项目的创新在于将合成粘附素作为 综合电路工程工具箱，并建立一个新的范式，为模块化工程的多细胞 生活材料。因此，该项目将广泛影响合成财团的工程， 研究，并使未来的健康应用的动态范围。

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.