Synthetic biology, with its goal of designing biological entities for wide-ranging purposes, remains a field of intensive research interest. However, the vast complexity of biological systems has heretofore rendered rational design prohibitively difficult. As a result, directed evolution remains a valuable tool for synthetic biology, enabling the identification of desired functionalities from large libraries of variants. This review highlights the most recent advances in the use of directed evolution in synthetic biology, focusing on new techniques and applications at the pathway and genome scale.
► Recent advances in directed evolution applied to synthetic biology are reviewed. ► New strategies facilitate library creation and in vivo evolution. ► Recent applications target biosynthetic and signal transduction pathways. ► New tools enable multiplex genome evolution.