Staufer Oskar, Schröter Martin, Platzman Ilia, Spatz Joachim P
Department for Cellular Biophysics, Max Planck Institute for Medical Research, Jahnstraße 29, 69120, Heidelberg, Germany.
Institute for Physical Chemistry, Department for Biophysical Chemistry, University of Heidelberg, Im Neuenheimer Feld 253, 69120, Heidelberg, Germany.
Small. 2020 Jul;16(27):e1906424. doi: 10.1002/smll.201906424. Epub 2020 Feb 20.
Bottom-up synthetic biology has directed most efforts toward the construction of artificial compartmentalized systems that recreate living cell functions in their mechanical, morphological, or metabolic characteristics. However, bottom-up synthetic biology also offers great potential to study subcellular structures like organelles. Because of their intricate and complex structure, these key elements of eukaryotic life forms remain poorly understood. Here, the controlled assembly of lipid enclosed, organelle-like architectures is explored by droplet-based microfluidics. Three types of giant unilamellar vesicles (GUVs)-based synthetic organelles (SOs) functioning within natural living cells are procedured: (A) synthetic peroxisomes supporting cellular stress-management, mimicking an organelle innate to the host cell by using analogous enzymatic modules; (B) synthetic endoplasmic reticulum (ER) as intracellular light-responsive calcium stores involved in intercellular calcium signalling, mimicking an organelle innate to the host cell but utilizing a fundamentally different mechanism; and (C) synthetic magnetosomes providing eukaryotic cells with a magnetotactic sense, mimicking an organelle that is not natural to the host cell but transplanting its functionality from other branches of the phylogenetic tree. Microfluidic assembly of functional SOs paves the way for high-throughput generation of versatile intracellular structures implantable into living cells. This in-droplet SO design may support or expand cellular functionalities in translational nanomedicine.
自下而上的合成生物学已将大部分精力投入到构建人工分隔系统中，这些系统在机械、形态或代谢特征方面重现活细胞的功能。然而，自下而上的合成生物学在研究细胞器等亚细胞结构方面也具有巨大潜力。由于其结构复杂精细，这些真核生物生命形式的关键元素仍未被充分理解。在此，通过基于液滴的微流控技术探索脂质包裹的、类似细胞器结构的可控组装。构建了三种在天然活细胞内发挥功能的基于巨型单层囊泡（GUVs）的合成细胞器（SOs）：（A）支持细胞应激管理的合成过氧化物酶体，通过使用类似的酶模块模拟宿主细胞固有的细胞器；（B）作为参与细胞间钙信号传导的细胞内光响应钙库的合成内质网（ER），模拟宿主细胞固有的细胞器，但利用了根本不同的机制；（C）为真核细胞提供趋磁感觉的合成磁小体，模拟一种宿主细胞非天然的细胞器，但从系统发育树的其他分支移植其功能。功能性SOs的微流控组装为高通量生成可植入活细胞的通用细胞内结构铺平了道路。这种液滴内SO设计可能会在转化纳米医学中支持或扩展细胞功能。
