Genome-wide CRISPRi-based identification of targets for decoupling growth from production.

Growth decoupling can be used to optimize microbial production of bio-based compounds by inhibiting excess biomass formation and re-direct carbon flux from growth to product formation. However, identifying suitable genetic targets through rational design is challenging. Here, we conduct a genome-wide CRISPRi screen to discover growth switches suitable for decoupling growth and production. Using an sgRNA library covering 12,238 loci in the genome, we screen for targets that inhibit growth while allowing for continued protein production. In total, we identify 1,332 sgRNAs that simultaneously decrease growth and maintains or increases accumulation of GFP. The top target / shows more than 5-fold increase in GFP accumulation and 45% decrease in biomass formation. Overall, our genome-wide CRISPRi screen provides key targets for growth decoupling, and the approach can be applied to improve bio-based production in other microorganisms.