Bottom-up synthetic biology thrives to reconstruct basic cellular processes into a minimalist cellular replica to foster their investigation in greater details with a reduced number of variables. Among these cellular features, the endomembrane system is an important aspect of cells which is at the origin of many of their functions. Still, the reconstruction of these inner compartments within a lipid-based vesicle remains challenging and poorly controlled. Herein, we report the use of pH as external trigger to self-assemble compartmentalized giant unilamellar vesicles (GUVs) by either bulk, or droplet-based microfluidics. By co-encapsulating pH sensitive small unilamellar vesicles (SUVs), negatively charged SUVs and/or proteins, we show that acidification of the droplets efficiently produces GUVs while sequestrating the co-encapsulated material with flexibility and robustness. The method enables the simultaneous reconstruction of more than a single cellular phenotype from the bottom-up, corresponding to an important advancement in the current status quo of bottom-up synthetic biology.

中文翻译：

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合成细胞中内膜多室的 pH 触发组装

自下而上的合成生物学蓬勃发展，将基本的细胞过程重建为极简主义的细胞复制品，以减少变量数量来促进对更详细的研究。在这些细胞特征中，内膜系统是细胞的一个重要方面，它是细胞许多功能的起源。尽管如此，在基于脂质的囊泡内重建这些内部隔室仍然具有挑战性且控制不佳。在此，我们报告了使用 pH 作为外部触发，通过散装或基于液滴的微流体自组装分隔的巨型单层囊泡 (GUV)。通过共同封装 pH 敏感的小单层囊泡 (SUV)、带负电荷的 SUV 和/或蛋白质，我们表明，液滴的酸化可有效地产生 GUV，同时以柔韧性和坚固性隔离共封装材料。该方法能够从自下而上同时重建多个细胞表型，这对应于自下而上合成生物学现状的重要进步。