A primer to directed evolution: current methodologies and future directions

Sellés Vidal, Lara; Isalan, Mark; Heap, John T.; Ledesma-Amaro, Rodrigo

Abstract

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.

Citation

Sellés Vidal, L., Isalan, M., Heap, J. T., & Ledesma-Amaro, R. (2023). A primer to directed evolution: current methodologies and future directions. RSC Chemical Biology, 2023(4), 271-291. https://doi.org/10.1039/d2cb00231k

• Journal Article Type: Article
• Acceptance Date: Jan 18, 2023
• Online Publication Date: Jan 27, 2023
• Publication Date: Apr 1, 2023
• Deposit Date: Feb 20, 2023
• Publicly Available Date: Feb 20, 2023
• Journal: RSC Chemical Biology
• Electronic ISSN: 2633-0679
• Publisher: Royal Society of Chemistry
• Peer Reviewed: Peer Reviewed
• Volume: 2023
• Issue: 4
• Pages: 271-291
• DOI: https://doi.org/10.1039/d2cb00231k
• Keywords: Biochemistry, Genetics and Molecular Biology (miscellaneous); Molecular Biology; Biochemistry; Chemistry (miscellaneous)
• Public URL: https://nottingham-repository.worktribe.com/output/16790055
• Publisher URL: https://pubs.rsc.org/en/content/articlelanding/2023/CB/D2CB00231K