Chinese hamster ovary (CHO) cells are widely used in the industrial production of biopharmaceuticals and it is the main industrial force of mammalian glycoprotein production. Many applications in CHO cells require the generation of stable cell lines and cultures lasting more than 1 week.
Instantaneous systems are becoming more and more popular. At present, various of genetic, chemical and environmental methods have been developed to regulate cellular pathways to improve titer. However, these methods are usually irreversible or have off target effect. The development of precise, tunable and reversible techniques will promote the time regulation of the target pathway to maximize the titer.
CRISPR-dCas9 system controlled by CRY2/CIB1 optogenetic switch can be used to regulate transient gene expression. This CHO expression system construction service based on optogenetic switch is now accessible at Creative BioMart. Our one-stop service covers all steps from GOI design to cell line delivery.
The activation of CHO transient system depends on the light activated dimerization of CRY2 and CIBN. CRY2 is a photosensitive protein, and its conformation changes when excited by 450 nm light. Here, CRY2 is fused with a viral transactivation domain to activate the expression of CMV promoter. CIBN and dCas9 are integrated into CRISPR-dCas-9 system. With the help of guide RNA (gRNA), the expression of target genes can be activated after photoactivated dimerization of CRY2 and CIBN. Once the light disappears, CRY2 and CIBN dissociate and gene expression is turned off.
| Step | Service Description |
| Expression strategy design | Client provides the target sequence and expression strategy designed referring to customers' requirement. |
| Plasmid design | Codon optimization Clone the GOI sequence downstream the CMV promoter |
| Plasmid transformation | Transfer the recombinant plasmid into the host cell |
| Light induction test | 450 nm light is used to stimulate the GOI expression to test the expression efficient. |
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.
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