The expanded CRISPR toolbox for constructing microbial cell factories

Yuxi Teng; Tian Jiang; Yajun Yan

doi:10.1016/j.tibtech.2023.06.012

The expanded CRISPR toolbox for constructing microbial cell factories

Trends Biotechnol. 2024 Jan;42(1):104-118. doi: 10.1016/j.tibtech.2023.06.012. Epub 2023 Jul 26.

Authors

Yuxi Teng¹, Tian Jiang¹, Yajun Yan²

Affiliations

¹ School of Chemical, Materials and Biomedical Engineering, College of Engineering, University of Georgia, Athens, GA 30602, USA.
² School of Chemical, Materials and Biomedical Engineering, College of Engineering, University of Georgia, Athens, GA 30602, USA. Electronic address: yajunyan@uga.edu.

PMID: 37500408
PMCID: PMC10808275 (available on 2025-01-01)
DOI: 10.1016/j.tibtech.2023.06.012

Abstract

Microbial cell factories (MCFs) convert low-cost carbon sources into valuable compounds. The CRISPR/Cas9 system has revolutionized MCF construction as a remarkable genome editing tool with unprecedented programmability. Recently, the CRISPR toolbox has been significantly expanded through the exploration of new CRISPR systems, the engineering of Cas effectors, and the incorporation of other effectors, enabling multi-level regulation and gene editing free of double-strand breaks. This expanded CRISPR toolbox powerfully promotes MCF construction by facilitating pathway construction, enzyme engineering, flux redistribution, and metabolic burden control. In this article, we summarize different CRISPR tool designs and their applications in MCF construction for gene editing, transcriptional regulation, and enzyme modulation. Finally, we also discuss future perspectives for the development and application of the CRISPR toolbox.

Keywords: CRISPR toolbox; base editing; metabolic engineering; microbial cell factory; synthetic metabolons; tunable regulation.

Publication types

Review

MeSH terms

CRISPR-Cas Systems*
Gene Editing*

Grants and funding

R35 GM128620/GM/NIGMS NIH HHS/United States