In vitro self-replication and multicistronic expression of large synthetic genomes
In vitro self-replication and multicistronic expression of large synthetic genomes
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DOI：
10.1038/s41467-020-14694-2
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发表时间：
2020-02-14
影响因子：
16.6
通讯作者：
Mutschler, H.
中科院分区：
文献类型：
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作者：
Libicher, K.;Hornberger, R.;Mutschler, H.
能够复制和进化的化学系统的产生是合成生物学的一个关键目标。这可以通过体外重建一个由DNA复制、转录和翻译组成的最小的自我维持的中心教条来实现。在这里，我们提出了一个体外翻译系统，它能够在定义明确的无细胞条件下自我编码复制和表达大的DNA基因组。特别是，我们展示了超过116kb的多部分基因组的自我复制，包括全套大肠杆菌翻译因子、所有三个核糖体RNA、能量再生系统以及RNA和DNA聚合酶。与DNA复制并行，我们的系统可以合成至少30个编码的翻译因子，其中一半的表达数量等于或大于它们各自的输入水平。我们优化的无细胞表达平台可以为建立部分自我复制的体外翻译系统提供基础。
The generation of a chemical system capable of replication and evolution is a key objective of synthetic biology. This could be achieved by in vitro reconstitution of a minimal self-sustaining central dogma consisting of DNA replication, transcription and translation. Here, we present an in vitro translation system, which enables self-encoded replication and expression of large DNA genomes under well-defined, cell-free conditions. In particular, we demonstrate self-replication of a multipartite genome of more than 116 kb encompassing the full set of Escherichia coli translation factors, all three ribosomal RNAs, an energy regeneration system, as well as RNA and DNA polymerases. Parallel to DNA replication, our system enables synthesis of at least 30 encoded translation factors, half of which are expressed in amounts equal to or greater than their respective input levels. Our optimized cell-free expression platform could provide a chassis for the generation of a partially self-replicating in vitro translation system.