Advanced research and exploration of CRISPR technology in the field of directed evolution

Rui Long; Dandan Tang; Tangli Yang; Mengyu Li; Yuting Lu; Wei Liu; Ling Jiang

doi:10.1016/j.biotechadv.2025.108633

Advanced research and exploration of CRISPR technology in the field of directed evolution

Biotechnol Adv. 2025 Jun 28:83:108633. doi: 10.1016/j.biotechadv.2025.108633. Online ahead of print.

Authors

Rui Long¹, Dandan Tang², Tangli Yang¹, Mengyu Li¹, Yuting Lu¹, Wei Liu³, Ling Jiang⁴

Affiliations

¹ College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China.
² School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211816, China.
³ College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China. Electronic address: liuwei6775@njtech.edu.cn.
⁴ College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China; State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing Tech University, Nanjing 211816, China. Electronic address: jiangling@njtech.edu.cn.

PMID: 40588042
DOI: 10.1016/j.biotechadv.2025.108633

Abstract

Directed molecular evolution is the key technology for obtaining enzymes, proteins, metabolic pathways, and other components of living organisms that have specific functions or desirable properties, which are indispensable in a variety of industrial and medical applications. Despite the success of traditional methods, they are often limited by low efficiency and the high cost of obtaining desired mutants. The advent of CRISPR technology has significantly advanced the field by enabling precise and efficient gene targeting, offering new prospects for directed evolution. This review provides a comprehensive overview of CRISPR tools and their applications in directed evolution, highlighting the principles, technological advancements, and specific applications of CRISPR-based mutation and screening platforms. We discuss the key findings from the use of CRISPR in enzyme and genome evolution, showcasing its ability to generate genetic diversity and select for improved phenotypes. The study underscores the unique value of CRISPR in directed evolution, particularly in its flexibility to target and edit various species' genomes, and its potential to accelerate the discovery of novel biomolecules with enhanced properties.

Keywords: CRISPR-Cas; Directed evolution; Enzyme evolution; Genome evolution.

Publication types

Review