A primer to directed evolution: current methodologies and future directions

Directed evolution is one of the most powerful tools for protein engineering and functions by harnessing natural evolution, but on a shorter timescale. It enables the rapid selection of variants of biomolecules with properties that make them more suitable for specific applications. Since the first in vitro evolution experiments performed by Sol Spiegelman in 1967, a wide range of techniques have been developed to tackle the main two steps of directed evolution: genetic diversification (library generation), and isolation of the variants of interest. This review covers the main modern methodologies, discussing the advantages and drawbacks of each, and hence the considerations for designing directed evolution experiments. Furthermore, the most recent developments are discussed, showing how advances in the handling of ever larger library sizes are enabling new research questions to be tackled.

Automated summary

Directed evolution is a powerful tool for protein engineering, allowing for the rapid selection of biomolecule variants with desired properties. Since its initial development in 1967, various techniques have been developed to facilitate genetic diversification and variant isolation. This review discusses the advantages, drawbacks, and recent developments in directed evolution methodologies, highlighting the ability to handle larger library sizes and address new research questions.