Optogenetic CRISPRi Platform
Microbial synthesis of chemicals has become an alternative method of chemical synthesis. However, the competition between natural metabolic pathways and heterologous pathways usually leads to metabolic imbalance and suboptimal production. In order to maximize cell productivity, it is necessary to balance metabolic pathways to obtain optimal strain performance.
Chemical Induction Vs. Light Induction
It is promising to interfere with the dynamic regulation of target gene expression by sensing specific signals, but there are still many limitations, such as the need for expensive inducers. In addition, inducers cannot be removed during fermentation, which makes it impossible for such systems to switch between activated and inactivated states, thus limiting their industrial application.
Optogenetic control of protein function and metabolic process can be realized by changing light conditions. Due to the advantages of time control and reversibility, optogenetic tools have been widely explored in the research related to neuroscience and tissue and cell development.
Optogenetic CRISPRi Solution at Creative BioMart
Creative BioMart optimized a blue light response suppression system based on EL222. When cells are exposed to blue light, the CRISPRi system is turned off to allow cell growth. In the dark, CRISPRi system inhibits the target gene, and carbon metabolic flux mainly turns to the synthesis of target protein. By adjusting the blue light frequency, the metabolic flux can be redistributed between cell growth and the synthesis of target products.
We provide construction services and protein expression services, including codon optimization, gene synthesis, protein expression and two-step purification. If you need other services, such as protein refolding, label removal and endotoxin removal services, you can also make additional choices. Through our proprietary codon optimization software and our experienced scientific team, we guarantee the delivery of high-quality final products.
| Service Name |
Service Description |
Timeline |
Deliverable |
| Gene synthesis and cloning, plasmid preparation |
- Codon optimization, gene synthesis, subcloning to expression vector
- Plasmid preparation
|
2-4 weeks |
Sequencing report |
| Protein expression evaluation |
- Transformation into proper strain
- Small-batch fermentation
- SDS-PAGE analysis to evaluate protein expression
|
1-2 weeks |
Expression evaluation result |
| Light induced protein expression |
- Photoinduced expression in E.coli
- Induce the protein expression with blue light
|
- |
|
| Protein expression and purification |
- Fermentation based on expression evaluation
- Protein purification
- Soluble expression-affinity purification
- Inclusion body expression-refolding
- SDS-PAGE and UV analysis
|
2-3 weeks |
- Feasibility report
- If expression is feasible, provide a certain amount of protein sample to the client based on the expression.
|
| Scale up and protein purification |
- Amplification Culture
- Preparation of crude extract
- Affinity purification
- Cut off fusion tag and remove protease (if necessary)
|
2-3 weeks |
- Purified protein
- SDS-PAGE report
|
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.
