Media Laboratory, Massachusetts Institute of Technology, Cambridge, MA, USA.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Nat Methods. 2022 Jan;19(1):55-64. doi: 10.1038/s41592-021-01348-4. Epub 2021 Dec 30.
Evolution occurs when selective pressures from the environment shape inherited variation over time. Within the laboratory, evolution is commonly used to engineer proteins and RNA, but experimental constraints have limited the ability to reproducibly and reliably explore factors such as population diversity, the timing of environmental changes and chance on outcomes. We developed a robotic system termed phage- and robotics-assisted near-continuous evolution (PRANCE) to comprehensively explore biomolecular evolution by performing phage-assisted continuous evolution in high-throughput. PRANCE implements an automated feedback control system that adjusts the stringency of selection in response to real-time measurements of each molecular activity. In evolving three distinct types of biomolecule, we find that evolution is reproducibly altered by both random chance and the historical pattern of environmental changes. This work improves the reliability of protein engineering and enables the systematic analysis of the historical, environmental and random factors governing biomolecular evolution.
当环境中的选择压力随时间塑造遗传变异时，进化就会发生。在实验室中，进化通常用于工程蛋白质和 RNA，但实验限制了可重复性和可靠性，无法探索种群多样性、环境变化的时机以及结果的偶然性等因素。我们开发了一种名为噬菌体和机器人辅助近连续进化（PRANCE）的机器人系统，通过高通量的噬菌体辅助连续进化来全面探索生物分子进化。PRANCE 实现了一个自动化反馈控制系统，该系统根据每个分子活性的实时测量来调整选择的严格程度。在进化三种不同类型的生物分子时，我们发现进化既可以被随机机会，也可以被环境变化的历史模式重复改变。这项工作提高了蛋白质工程的可靠性，并使系统分析控制生物分子进化的历史、环境和随机因素成为可能。
