Publication Date
7-17-2023
Journal
Nature Communications
Abstract
Synthetic biology requires efficient systems that support the well-coordinated co-expression of multiple genes. Here, we discover a 9-bp nucleotide sequence that enables efficient polycistronic gene expression in yeasts and filamentous fungi. Coupling polycistronic expression to multiplexed, markerless, CRISPR/Cas9-based genome editing, we develop a strategy termed HACKing (Highly efficient and Accessible system by CracKing genes into the genome) for the assembly of multigene pathways. HACKing allows the expression level of each enzyme to be precalibrated by linking their translation to those of host proteins with predetermined abundances under the desired fermentation conditions. We validate HACKing by rapidly constructing highly efficient Saccharomyces cerevisiae cell factories that express 13 biosynthetic genes, and produce model endogenous (1,090.41 ± 80.92 mg L
Keywords
CRISPR-Cas Systems, Gene Editing, Saccharomyces cerevisiae, Fungi
Included in
Bioinformatics Commons, Biomedical Informatics Commons, Fungi Commons, Medical Sciences Commons, Medical Specialties Commons
Comments
Supplementary Materials
PMID: 37460548