Biosensor-guided genome-wide CRISPR screens enable cis,cis-muconic acid overproduction in yeast

In Saccharomyces cerevisiae, biosensor-aided MAGIC (senMAGIC) identified single-gene perturbations that increase cis,cis-muconic acid titres, including novel targets confirmed by genomic integration.

• Why it matters: Classical metabolic engineering is slow and can miss unintuitive genetic changes; genome-wide CRISPR screens need high-throughput readouts to efficiently enrich improved strains.
• What they did: The team built small-molecule biosensors in yeast using zeocin resistance and fluorescence, then ran a MAGIC library targeting every gene for activation, interference, or deletion (senMAGIC). They used senMAGIC to find determinants for protocatechuic acid and cis,cis-muconic acid, then tested combinatorial effects with triple gRNA-tRNA arrays and confirmed top hits via direct genomic integrations in senMAGIC-free strains.
• The result: The confirmed perturbations increased PCA and ccM titres and revealed both known and novel targets, supporting senMAGIC as an automation-friendly whole-genome evolution approach while outlining method limitations.

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