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Related Experiment Video

Updated: Jun 2, 2026

A Practical Guide to Phage- and Robotics-Assisted Near-Continuous Evolution

A Practical Guide to Phage- and Robotics-Assisted Near-Continuous Evolution

Published on: January 12, 2024

A system for the continuous directed evolution of biomolecules.

Kevin M Esvelt¹, Jacob C Carlson, David R Liu

¹Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.

|April 12, 2011

Summary

Phage-assisted continuous evolution (PACE) enables rapid, continuous laboratory evolution of gene-encoded molecules in bacteria. This breakthrough accelerates the discovery of novel biomolecules and enzymes, significantly enhancing evolutionary effectiveness.

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Area of Science:

Molecular Biology
Biotechnology
Evolutionary Biology

Background:

Laboratory evolution accelerates biomolecule development but is slow and labor-intensive.
Previous continuous evolution methods were limited to specific molecules like ribozymes.
Accelerating evolutionary cycles is key to enhancing biomolecule engineering.

Purpose of the Study:

To develop a system for continuous directed evolution of gene-encoded molecules in bacteria.
To significantly increase the speed and efficiency of laboratory evolution.
To enable rapid engineering of proteins with novel functions.

Main Methods:

Phage-assisted continuous evolution (PACE) system utilizing a modified bacteriophage life cycle in Escherichia coli.
Genes are transferred between host cells dependent on the desired molecular activity.
PACE allows for dozens of evolution rounds per day without human intervention.

Main Results:

PACE successfully evolved T7 RNA polymerase (RNAP) variants with altered promoter specificity and nucleotide initiation.
Engineered RNAP variants showed hundreds-fold improvements in activity compared to wild-type.
200 rounds of evolution were completed in 8 days, yielding enzymes with desired activities from undetectable levels.

Conclusions:

PACE dramatically accelerates laboratory evolution, enabling rapid discovery of functional biomolecules.
The system overcomes limitations of previous directed evolution techniques.
PACE offers a powerful tool for tackling complex protein engineering challenges and studying molecular evolution.