Simultaneous On and Off Gene Control with Gene Scissors (IMAGE)
Caption
< (Left) The principle of the dual-mode CRISPR gene scissors. When the guide RNA (gRNA) binds to the target sequence, dxCas9-CRP either promotes (CRISPRa) or inhibits (CRISPRi) the binding of RNA polymerase near the transcription start site, precisely controlling gene expression. (Center) A large-scale screening of the entire E. coli genome is conducted to identify key regulatory targets for optimizing target substance production. The metabolic pathway for producing the target substance is then re-engineered by simultaneously regulating gene expression through activation and repression. (Right) The dual-mode CRISPR gene scissors system enables systematic redesign of cell metabolism, precise reconfiguration of gene expression, and the construction of microbial strains that can perform various functions, ultimately leading to a significant increase in target substance productivity. In this study, the dual-mode CRISPR system was applied to E. coli to demonstrate the enhanced production of 'violacein,' a purple functional biopigment with anticancer effects, and its potential for expansion to other bacterial species was also confirmed. >
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