Gene Expression in on-Chip Membrane-Bound Artificial Cells
Gene Expression in on-Chip Membrane-Bound Artificial Cells
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DOI：
10.1021/acssynbio.9b00247
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发表时间：
2019-08-01
影响因子：
4.7
通讯作者：
Maeda, Yusuke T.
中科院分区：
文献类型：
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作者：
Izri, Ziane;Garenne, David;Maeda, Yusuke T.
由分子组分和脂质膜组成的人工细胞是表征生命系统特性的新兴平台。无细胞转录-翻译（TXTL）为细胞反应器的自下而上合成提供了优势。然而，在定义明确的几何形状内扩展其设计仍然具有挑战性。我们提出了一种微流控装置托管TXTL反应的报告基因在数千个微孔分离的外部缓冲液的磷脂膜。在缓冲液中存在营养物的情况下，微反应器稳定超过24小时，并产生几mg/mL的蛋白质。尽管没有孔，外部溶液中的营养物经由穿过每个微流体孔的磷脂膜的被动运输以皮升规模供给TXTL反应。用等渗浓度的惰性聚合物和脂肪酸代替营养物会降低微反应器的效率，并且很大一部分不会产生蛋白质。这强调了膜在设计无细胞TXTL微反应器作为高效人工细胞中的关键作用。
Artificial cells made of molecular components and lipid membrane are emerging platforms to characterize living systems properties. Cell-free transcription-translation (TXTL) offers advantages for the bottom-up synthesis of cellular reactors. Yet, scaling up their design within well-defined geometries remains challenging. We present a microfluidic device hosting TXTL reactions of a reporter gene in thousands of microwells separated from an external buffer by a phospholipid membrane. In the presence of nutrients in the buffer, microreactors are stable beyond 24 h and yield a few mg/mL of proteins. Nutrients in the external solution feed the TXTL reaction at the picoliter scale via passive transport across the phospholipid membrane of each microfluidic well, despite the absence of pores. Replacing nutrients with an inert polymer and fatty acids at an isotonic concentration reduces microreactors efficiency, and a significant fraction yields no protein. This emphasizes the crucial role of the membrane for designing cell-free TXTL microreactors as efficient artificial cells.