Issue 4, 2023

A primer to directed evolution: current methodologies and future directions

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.

Graphical abstract: A primer to directed evolution: current methodologies and future directions

Article information

Article type
Review Article
Submitted
21 Nov 2022
Accepted
18 Jan 2023
First published
27 Jan 2023
This article is Open Access
Creative Commons BY license

RSC Chem. Biol., 2023,4, 271-291

A primer to directed evolution: current methodologies and future directions

L. Sellés Vidal, M. Isalan, J. T. Heap and R. Ledesma-Amaro, RSC Chem. Biol., 2023, 4, 271 DOI: 10.1039/D2CB00231K

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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