Journal
ACS SYNTHETIC BIOLOGY
Volume 9, Issue 5, Pages 1030-1040Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.9b00143
Keywords
CRISPRi; growth decoupling; bioproduction; genome-wide screening; E. coli; growth inhibition
Categories
Funding
- People Programme (Marie Curie Actions) of the European Union Seventh Framework Programme FP7-People-2012-ITN [317058]
- Novo Nordisk Foundaion [NNF16CC0020908, NNF15OC0015246]
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Growth decoupling can be used to optimize microbial production of biobased compounds by inhibiting excess biomass formation and redirect 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 Escherichia coli genome, we screen for targets that inhibit growth while allowing for continued protein production. In total, we identify 1332 sgRNAs that simultaneously decrease growth and maintain or increase accumulation of GFP. The top target sibB/ibsB 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 biobased production in other microorganisms.
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