TY  - JOUR
A1  - Naseri, Gita
A1  - Behrend, Jessica
A1  - Rieper, Lisa
A1  - Müller-Röber, Bernd
T1  - COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors
JF  - Nature Communications
N2  - 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 n-carotene and co-producing p-ionone and biosensor-responsive naringenin. COMPASS will have many applications in synthetic biology projects that require gene expression balancing.
Y1  - 2019
U6  - https://doi.org/10.1038/s41467-019-10224-x
SN  - 2041-1723
VL  - 10
PB  - Nature Publ. Group
CY  - London
ER  -