Automated Continuous Evolution of Proteins in Vivo

Area of Science:

• Biotechnology and Synthetic Biology
• Molecular Biology
• Evolutionary Biology

Background:

• Directed evolution is crucial for protein engineering but often requires extensive manual labor.
• Existing methods for biomolecular evolution can be time-consuming and lack scalability.
• Automated platforms are needed to accelerate the discovery of novel biomolecular functions.

Purpose of the Study:

• To introduce Automated Continuous Evolution (ACE), a novel platform for hands-free, directed evolution of biomolecules.
• To demonstrate the capability of ACE in achieving desired functions such as drug resistance and enhanced enzyme activity.
• To highlight the potential of ACE for protein engineering and evolutionary studies.

Main Methods:

• ACE integrates OrthoRep (in vivo targeted mutagenesis) with eVOLVER (automated continuous culture).
• Real-time feedback control of selection stringency was implemented using eVOLVER.
• Genes of interest were autonomously evolved through multimutation adaptive pathways.

Main Results:

• ACE successfully enabled hands-free, directed evolution of biomolecules.
• The platform facilitated autonomous traversal of adaptive pathways leading to desired functions.
• Achieved outcomes included enhanced drug resistance and improved enzyme activity.

Conclusions:

• ACE offers a durable, scalable, and rapid solution for biomolecular evolution.
• The platform has broad applicability in protein engineering and evolutionary adaptation studies.
• ACE facilitates investigations into how selection parameters influence evolutionary trajectories.