Gene

Banach space

Pure mathematics

Biology

Computer science

Molecular evolution

Computational biology

Genetics

Phylogenetic tree

Mathematics

Mutant

Sequence space

Directed Molecular Evolution

Directed evolution

Nature Chemical Biology

Continuous directed evolution methods allow the key steps of evolution—gene diversification, selection, and replication—to proceed in the laboratory with minimal researcher intervention. As a result, continuous evolution can find solutions much more quickly than traditional discrete evolution methods. Continuous evolution also enables the exploration of longer and more numerous evolutionary trajectories, increasing the likelihood of accessing solutions that require many steps through sequence space and greatly facilitating the iterative refinement of selection conditions and targeted mutagenesis strategies. Here we review the historical advances that have expanded continuous evolution from its earliest days as an experimental curiosity to its present state as a powerful and surprisingly general strategy for generating tailor-made biomolecules, and discuss more recent improvements with an eye to the future. This historical Perspective on continuous directed evolution focuses on laboratory approaches that enable greater understanding of evolving molecular populations and offer investigators tools to guide the emergence of new biomolecular systems.

The developing toolkit of continuous directed evolution