One of the aims of synthetic biology is bottom-up construction of reconstituted human cells for medical uses. To that end, we generated giant unilamellar vesicles (GUVs) that contained a HeLa cell extract, which comprises a cell-free protein synthesis (CFPS) system. Then we expressed Huntingtin protein fragments that contained polyglutamine (polyQ) sequences (Htt-polyQ), a hallmark of Huntington’s disease. That system produced polyQ-dependent protein aggregates, as previously demonstrated in living cells. We next simplified the system by generating GUVs that contained purified human factors, which reconstituted a CFPS system. Htt-polyQ fragments expressed in these GUVs also formed protein aggregates. Moreover, an N-terminal deletion mutant, which had failed to form protein aggregates in living cells, also failed to form protein aggregates in the reconstituted GUVs. Thus, the GUV systems that encapsulated a human CFPS system could serve as reconstituted cells for studying neurological diseases.

中文翻译：

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在重构细胞中Huntingtin聚谷氨酰胺依赖性蛋白聚集

合成生物学的目的之一是自下而上构建用于医学用途的重组人细胞。为此，我们生成了包含HeLa细胞提取物的巨型单层囊泡（GUV），该提取物包含无细胞蛋白质合成（CFPS）系统。然后，我们表达了包含亨廷顿氏病标志的聚谷氨酰胺（polyQ）序列（Htt-polyQ）的亨廷顿蛋白片段。该系统产生了polyQ依赖的蛋白质聚集体，如先前在活细胞中所证明的。接下来，我们通过生成包含纯化的人为因素的GUV来简化系统，从而重构了CFPS系统。这些GUV中表达的Htt-polyQ片段也形成蛋白质聚集体。而且，N-末端缺失突变体未能在活细胞中形成蛋白质聚集体，也未能在重构的GUV中形成蛋白质聚集体。因此，封装了人CFPS系统的GUV系统可以用作研究神经系统疾病的重构细胞。