TY  - GEN
A1  - Messerschmidt, Katrin
A1  - Machens, Fabian
A1  - Hochrein, Lena
A1  - Naseri, Gita
T1  - Orthogonal, light-inducible protein expression platform in yeast Sacchararomyces cerevisiae
T2  - New biotechnology
Y1  - 2018
U6  - https://doi.org/10.1016/j.nbt.2018.05.153
SN  - 1871-6784
SN  - 1876-4347
VL  - 44
SP  - S19
EP  - S19
PB  - Elsevier
CY  - Amsterdam
ER  - 
TY  - JOUR
A1  - Naseri, Gita
A1  - Balazadeh, Salma
A1  - Machens, Fabian
A1  - Kamranfar, Iman
A1  - Messerschmidt, Katrin
A1  - Müller-Röber, Bernd
T1  - Plant-Derived Transcription Factors for Orthologous Regulation of Gene Expression in the Yeast Saccharomyces cerevisiae
JF  - ACS synthetic biology
N2  - Control of gene expression by transcription factors (TFs) is central in many synthetic biology projects for which a tailored expression of one or multiple genes is often needed. As TFs from evolutionary distant organisms are unlikely to affect gene expression in a host of choice, they represent excellent candidates for establishing orthogonal control systems. To establish orthogonal regulators for use in yeast (Saccharomyces cerevisiae), we chose TFs from the plant Arabidopsis thaliana. We established a library of 106 different combinations of chromosomally integrated TFs, activation domains (yeast GAL4 AD, herpes simplex virus VP64, and plant EDLL) and synthetic promoters harboring cognate cis regulatory motifs driving a yEGFP reporter. Transcriptional output of the different driver/reporter combinations varied over a wide spectrum, with EDLL being a considerably stronger transcription activation domain in yeast than the GAL4 activation domain, in particular when fused to Arabidopsis NAC TFs. Notably, the strength of several NAC-EDLL fusions exceeded that of the strong yeast TDH3 promoter by 6- to 10-fold. We furthermore show that plant TFs can be used to build regulatory systems encoded by centromeric or episomal plasmids. Our library of TF-DNA binding site combinations offers an excellent tool for diverse synthetic biology applications in yeast.
KW  - Arabidopsis thaliana
KW  - artificial transcription factor
KW  - NAC transcription factor
KW  - synthetic biology
KW  - plant
Y1  - 2017
U6  - https://doi.org/10.1021/acssynbio.7b00094
SN  - 2161-5063
VL  - 6
SP  - 1742
EP  - 1756
PB  - American Chemical Society
CY  - Washington
ER  - 
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  -