Zhanar Abil, Ana María Restrepo Sierra, Andreea R. Stan, Amélie Châne, Alicia del Prado, Miguel de Vega, Yannick Rondelez, Christophe Danelon
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引用次数: 0
Abstract
Replication, heredity, and evolution are characteristic of Life. We and others have postulated that the reconstruction of a synthetic living system in the laboratory will be contingent on the development of a genetic self-replicator capable of undergoing Darwinian evolution. Although DNA-based life dominates, the in vitro reconstitution of an evolving DNA self-replicator has remained challenging. We hereby emulate in liposome compartments the principles according to which life propagates information and evolves. Using two different experimental configurations supporting intermittent or semi-continuous evolution (i.e., with or without DNA extraction, PCR, and re-encapsulation), we demonstrate sustainable replication of a linear DNA template – encoding the DNA polymerase and terminal protein from the Phi29 bacteriophage – expressed in the ‘protein synthesis using recombinant elements’ (PURE) system. The self-replicator can survive across multiple rounds of replication-coupled transcription-translation reactions in liposomes and, within only ten evolution rounds, accumulates mutations conferring a selection advantage. Combined data from next-generation sequencing with reverse engineering of some of the enriched mutations reveal nontrivial and context-dependent effects of the introduced mutations. The present results are foundational to build up genetic complexity in an evolving synthetic cell, as well as to study evolutionary processes in a minimal cell-free system.
复制、遗传和进化是生命的特征。我们和其他人推测,要在实验室中重建合成生命系统,就必须开发出能够经历达尔文进化的基因自我复制器。虽然以 DNA 为基础的生命占主导地位,但在体外重建一个不断进化的 DNA 自我复制器仍然具有挑战性。因此,我们在脂质体中模拟生命传播信息和进化的原理。利用支持间歇性或半连续性进化的两种不同实验配置(即有无 DNA 提取、PCR 和再封装),我们展示了线性 DNA 模板的可持续复制,该模板编码 Phi29 噬菌体的 DNA 聚合酶和末端蛋白,并在 "利用重组元件进行蛋白质合成"(PURE)系统中表达。自我复制者可以在脂质体中经过多轮复制耦合转录-翻译反应后存活下来,并在短短十轮进化过程中积累突变,从而获得选择优势。下一代测序数据与部分富集突变的逆向工程相结合,揭示了引入的突变所产生的非同小可且与上下文相关的影响。本研究成果对于在不断进化的合成细胞中建立遗传复杂性以及研究最小无细胞系统中的进化过程具有奠基性意义。
期刊介绍:
Nature Communications, an open-access journal, publishes high-quality research spanning all areas of the natural sciences. Papers featured in the journal showcase significant advances relevant to specialists in each respective field. With a 2-year impact factor of 16.6 (2022) and a median time of 8 days from submission to the first editorial decision, Nature Communications is committed to rapid dissemination of research findings. As a multidisciplinary journal, it welcomes contributions from biological, health, physical, chemical, Earth, social, mathematical, applied, and engineering sciences, aiming to highlight important breakthroughs within each domain.