CRISPR-derived genome editing technologies for metabolic engineering.
作者： Keiji Nishida , Akihiko Kondo
DOI: 10.1016/J.YMBEN.2020.12.002
关键词:
摘要: In metabolic engineering, genome editing tools make it much easier to discover and evaluate relevant genes pathways construct strains. Clustered regularly interspaced palindromic repeats (CRISPR)-associated (Cas) systems now have become the first choice for engineering in many organisms includingindustrially ones. Targeted DNA cleavage by CRISPR-Cas provides variousgenome modes such as indels, replacements, large deletions, knock-in chromosomal rearrangements, while host-dependent differences repair need be considered. The versatility of CRISPR system has given rise derivative technologies that complement nuclease-based editing, which causes cytotoxicity especially microorganisms. Deaminase-mediated base installs targeted point mutations with less toxicity. CRISPRi CRISPRa can temporarily control gene expression without changing genomic sequence. Multiplex, combinatorial scale are made possible streamlined design construction gRNA libraries further accelerates comprehensive discovery, evaluation building pathways. This review summarizes technical basis recent advances CRISPR-related applied purposes, representative examples industrially eukaryotic prokaryotic organisms.
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