Budding yeast Saccharomyces cerevisiae has become the primary eukaryotic model organism in the study of cell and molecular biology. Synthetic biology has adopted yeast as a biological platform for different purposes including DNA assembly in vivo, production of high-value metabolites therapeutic protein. As a microbial cell factory, yeast can be cultured in low-cost medium. It has the characteristics of fast growth, high protein yield, target protein secretion and post-translational modification.

Creative BioMart has been engaged in microbial gene editing for many years and has a perfect Saccharomyces cerevisiae gene editing technology platform. Our gene editing technology platform has experienced technicians, efficient experimental process and perfect experimental equipment. Our scientists can customize one-to-one solutions according to the specific needs of customers, and provide editing services for a variety of microorganisms to meet the needs of customers.

Construction of the CRISPR-Cas9 +CRY 2 Yeast System

Creative BioMart combines photogenetic switch and CRISPR-Cas9 technology to insert photoreceptors and their interacting proteins into the transcriptional activation element of the target gene. An inactive version of Cas9 (dCas9) was used to replace the DNA binding domain (DBD), in which the C-terminal of dCas9 was fused with the cryptochrome interacting protein CIB1. The combination of crispr-cas9 and photogenetic switching technology extends the characteristics and advantages of light as a gene expression inducer, making it possible to specifically target any promoter sequence in the genome.

Recombinant Protein Expression System

Step	Description	Deliverable	Time
Subcloning of yeast expression vector	Codon optimization, gene synthesis (or customer provided plasmid template: target gene amplification) The target gene was subcloned into a suitable expression plasmid Splice sequence validation: sequencing	Correctly constructed recombinant expression plasmid, DH5 containing recombinant plasmid α Strain and sequencing original report	1-2 weeks
Yeast expressing strain transformation	Amplify and culture 5-10 positive clones and induce the expression of recombinant protein	Report of positive clone strains and expression screening results.	2 weeks
Screening of yeast expressing strains	Detection of recombinant protein expression (applicable to multi copy number and high expression strains)	Report of positive clone strains and expression screening results.	2 weeks
Optimization of yeast expression strains	For the selected monoclonal strains, the culture and induction conditions (medium, cell density, methanol concentration, induction time, etc.) were optimized to improve the expression of the target protein		2-3 weeks

Blue light controlled protein expression has the following advantages

• The transmission of light is immediate and does not require time to absorb chemicals into cells
• Light does not need to operate the medium, and can be easily automated
• Light is minimally invasive
• Light is not affected by compound absorption or drug pump problems
• The light induction system allows precise dose-dependent control of protein levels because light can be transmitted within a specified time and removed immediately

Creative BioMart is always devote us to provide high-quality and satisfactory service to our customers, if you are interested in our services or have some question, please feel free to contact us or make online inquiry.

Online inquiry