The Louis Family Laboratory for Targeted Drug Delivery and Personalized Medicine Technologies, Department of Chemical Engineering, Technion, Haifa, Israel.
The Norman Seiden Multidisciplinary Program for Nanoscience and Nanotechnology, Technion, Haifa, Israel.
Nat Commun. 2022 Apr 28;13(1):2328. doi: 10.1038/s41467-022-29871-8.
Development of regulated cellular processes and signaling methods in synthetic cells is essential for their integration with living materials. Light is an attractive tool to achieve this, but the limited penetration depth into tissue of visible light restricts its usability for in-vivo applications. Here, we describe the design and implementation of bioluminescent intercellular and intracellular signaling mechanisms in synthetic cells, dismissing the need for an external light source. First, we engineer light generating SCs with an optimized lipid membrane and internal composition, to maximize luciferase expression levels and enable high-intensity emission. Next, we show these cells' capacity to trigger bioprocesses in natural cells by initiating asexual sporulation of dark-grown mycelial cells of the fungus Trichoderma atroviride. Finally, we demonstrate regulated transcription and membrane recruitment in synthetic cells using bioluminescent intracellular signaling with self-activating fusion proteins. These functionalities pave the way for deploying synthetic cells as embeddable microscale light sources that are capable of controlling engineered processes inside tissues.
在合成细胞中开发受调控的细胞过程和信号传递方法对于它们与活体材料的整合至关重要。光作为一种有吸引力的工具可以实现这一点，但可见光在组织中的有限穿透深度限制了其在体内应用中的可用性。在这里，我们描述了在合成细胞中设计和实施生物发光细胞间和细胞内信号传递机制，无需外部光源。首先，我们通过优化脂质膜和内部成分来设计发光的 SCs，以最大限度地提高荧光素酶的表达水平并实现高强度发射。接下来，我们展示了这些细胞通过启动真菌深绿木霉的无性孢子形成来触发天然细胞中生物过程的能力。最后，我们使用具有自激活融合蛋白的生物发光细胞内信号传递来演示合成细胞中的转录和膜募集的调控。这些功能为将合成细胞用作可嵌入微尺度光源铺平了道路，这些光源能够控制组织内的工程化过程。
