Engineer biomolecules by linking desired properties with phage propagation

Area of Science:

• Molecular Biology
• Biotechnology
• Evolutionary Biology

Background:

• Genetic diversity is crucial for laboratory evolution, often achieved through mutagenesis and artificial selection.
• Continuous evolution integrates laboratory evolution steps into a cycle with minimal human intervention.
• Bacteriophages offer a robust platform for in vivo continuous evolution due to their mutation rates and manipulability.

Purpose of the Study:

• To introduce an innovative system for the directed evolution of biomolecules.
• To demonstrate the efficiency and broad applicability of phage-assisted continuous evolution (PACE).

Main Methods:

• Development of the phage-assisted continuous evolution (PACE) system.
• Application of PACE for optimizing various biomolecules, including RNAP, protease, gene editing tools, and antibody fragments.
• Creation of simplified PACE versions (PRECEL, PANCE) and a novel M13 phagemid-based system.

Main Results:

• PACE allows over 40 rounds of evolution per day, drastically reducing evolution time.
• Optimized biomolecules, such as RNAP and single-chain variable fragments (scFv), exhibit enhanced activities and specificities.
• Development of complementary systems (PRECEL, PANCE) expands the directed evolution toolkit.

Conclusions:

• Phage-assisted systems like PACE offer a powerful and rapid method for biomolecule evolution.
• These systems hold significant promise for optimizing disease-related molecules for improved biological therapies.
• The technology facilitates the development of more potent therapeutic agents for clinical application.