Journal
METABOLIC ENGINEERING
Volume 63, Issue -, Pages 141-147Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2020.12.002
Keywords
Genome editing; Metabolic engineering; CRISPR; Recombineering; Base editing
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Funding
- New Energy and Industrial Technology Development Organization (NEDO)
- Japan Agency for Medical Research and Development (AMED)
- Special Coordination Fund for Promoting Science and Technology
- Creation of Innovative Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe) from the Ministry of Education, Culture, Sports and Technology (MEXT) of Japan
- Cross-ministerial Strategic Innovation Promotion Program
- JSPS KAKENHI [26119710, 16K14654]
- Grants-in-Aid for Scientific Research [16K14654, 26119710] Funding Source: KAKEN
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CRISPR-Cas system offers various modes for DNA editing in metabolic engineering, with consideration of host-dependent repair pathways. The versatility of CRISPR system has led to derivative technologies to complement nuclease-based editing, avoiding cytotoxicity, especially in microorganisms.
In metabolic engineering, genome editing tools make it much easier to discover and evaluate relevant genes and pathways and construct strains. Clustered regularly interspaced palindromic repeats (CRISPR)-associated (Cas) systems now have become the first choice for genome engineering in many organisms includingindustrially relevant ones. Targeted DNA cleavage by CRISPR-Cas provides variousgenome engineering modes such as indels, replacements, large deletions, knock-in and chromosomal rearrangements, while host-dependent differences in repair pathways need to be considered. The versatility of the CRISPR system has given rise to derivative technologies that complement nuclease-based editing, which causes cytotoxicity especially in microorganisms. Deaminase-mediated base editing installs targeted point mutations with much less toxicity. CRISPRi and CRISPRa can temporarily control gene expression without changing the genomic sequence. Multiplex, combinatorial and large scale editing are made possible by streamlined design and construction of gRNA libraries to further accelerates comprehensive discovery, evaluation and building of metabolic pathways. This review summarizes the technical basis and recent advances in CRISPR-related genome editing tools applied for metabolic engineering purposes, with representative examples of industrially relevant eukaryotic and prokaryotic organisms.
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