High-throughput metabolic engineering of <i>Yarrowia lipolytica</i> through gene expression tuning.

ABSTRACT: The challenge of accurately predicting which genetic alternations lead to the desired phenotype necessitates high-throughput metabolic engineering approaches where numerous hypotheses can be tested simultaneously. We describe the CRISPR-Cas9-based method TUNE^YALI that enables high-throughput tuning of gene expression in the common industrial yeast Yarrowia lipolytica. The method is based on replacing the promoters of the target genes with native Y. lipolytica promoters of varying strengths or removing the promoters entirely. To demonstrate the method's capabilities, we created a plasmid library that targets 56 transcription factors (TFs) and changes the expression of each TF to seven different levels. We transformed this library into reference and betanin-producing

SUBMITTER: Jiang W 

PROVIDER: S-EPMC12168020 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature