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COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors.

ABSTRACT: Balanced expression of multiple genes is central for establishing new biosynthetic pathways or multiprotein cellular complexes. Methods for efficient combinatorial assembly of regulatory sequences (promoters) and protein coding sequences are therefore highly wanted. Here, we report a high-throughput cloning method, called COMPASS for COMbinatorial Pathway ASSembly, for the balanced expression of multiple genes in Saccharomyces cerevisiae. COMPASS employs orthogonal, plant-derived artificial transcription factors (ATFs) and homologous recombination-based cloning for the generation of thousands of individual DNA constructs in parallel. The method relies on a positive selection of correctly assembled pathway variants from both, in vivo and in vitro cloning procedures. To decrease the turnaround time in genomic engineering, COMPASS is equipped with multi-locus CRISPR/Cas9-mediated modification capacity. We demonstrate the application of COMPASS by generating cell libraries producing ?-carotene and co-producing ?-ionone and biosensor-responsive naringenin. COMPASS will have many applications in synthetic biology projects that require gene expression balancing.

SUBMITTER: Naseri G

PROVIDER: S-EPMC6565718 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors.

Naseri Gita G Behrend Jessica J Rieper Lisa L Mueller-Roeber Bernd B

Nature communications 20190613 1

Balanced expression of multiple genes is central for establishing new biosynthetic pathways or multiprotein cellular complexes. Methods for efficient combinatorial assembly of regulatory sequences (promoters) and protein coding sequences are therefore highly wanted. Here, we report a high-throughput cloning method, called COMPASS for COMbinatorial Pathway ASSembly, for the balanced expression of multiple genes in Saccharomyces cerevisiae. COMPASS employs orthogonal, plant-derived artificial tran ...[more]

PMID: 31197154

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Dataset Information

COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors.

Publications

COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors.

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