Biomimetic Systems, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152, Martinsried, Germany.
Intelligent Biointegrative Systems Group, University of Stuttgart, Pfaffenwaldring 57, 70569, Stuttgart, Germany.
Nat Commun. 2020 Feb 14;11(1):904. doi: 10.1038/s41467-020-14694-2.
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.
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中心法则来实现。在这里，我们提出了一个体外翻译系统，该系统能够在明确的无细胞条件下自我编码复制和表达大型 DNA 基因组。特别是，我们证明了一个超过 116kb 的多部分基因组的自我复制，该基因组包含了大肠杆菌翻译因子的全套、所有三个核糖体 RNA、一个能量再生系统，以及 RNA 和 DNA 聚合酶。与 DNA 复制平行，我们的系统能够合成至少 30 种编码的翻译因子，其中一半的表达量等于或大于其各自的输入水平。我们优化的无细胞表达平台可以为部分自我复制的体外翻译系统的生成提供一个底盘。
